David Tollervey

An Endoribonuclease Functionally Linked to Perinuclear mRNP Quality Control Associates with the Nuclear Pore Complexes (2009)

Michal Skružný, Claudia Schneider, Attila Rácz, Julan Weng, David Tollervey, Ed Hurt

Nuclear mRNA export is a crucial step in eukaryotic gene expression, which is in yeast coupled to cotranscriptional messenger ribonucleoprotein particle (mRNP) assembly and surveillance. Several...

The N-terminal PIN domain of the exosome subunit Rrp44 harbors endonuclease activity and tethers Rrp44 to the yeast core exosome (2009)

Schneider, Claudia, Leung, Eileen, Brown, Jeremy, Tollervey, David

Nuclear and cytoplasmic forms of the yeast exosome share 10 components, of which only Rrp44/Dis3 is believed to possess 3′ exonuclease activity. We report that expression only of Rrp44 lacking...

A yeast exosome cofactor, Mpp6, functions in RNA surveillance and in the degradation of noncoding RNA transcripts (2008)

Decourty, Laurence, Saveanu, Cosmin, Rappsilber, Juri, Jacquier, Alain, ...

A genome-wide screen for synthetic lethal (SL) interactions with loss of the nuclear exosome cofactors Rrp47/Lrp1 or Air1 identified 3' --> 35' exonucleases, the THO complex required for mRNP...

Modelling yeast pre-rRNA processing (2008)

Federica Ciocchetta, Jane Hillston, Martin Kos, David Tollervey

Abstract. In this paper we present a quantified model concerning the synthesis of pre-rRNAs. The chemical kinetics simulation software Dizzy has been chosen as both the modelling and simulation...

The mRNA encoding the yeast ARE-binding protein Cth2 is generated by a novel 3' processing pathway (2008)

Ciais, Delphine, Bohnsack, Markus T., Tollervey, David

Microarray analyses of mRNAs over-expressed in strains lacking the nuclear exosome component Rrp6 identified the transcript encoding the ARE-binding protein Cth2, which functions in cytoplasmic mRNA...

The yeast ribosome synthesis factor Emg1 is a novel member of the superfamily of alpha/beta knot fold methyltransferases (2008)

Leulliot, Nicolas, Bohnsack, Markus T., Graille, Marc, Tollervey, David, Van Tilbeurgh, Herman

Emg1 was previously shown to be required for maturation of the 18S rRNA and biogenesis of the 40S ribosomal subunit. Here we report the determination of the crystal structure of Emg1 at 2 Å...

Efficient termination of transcription by RNA polymerase I requires the 5' exonuclease Rat1 in yeast (2008)

El Hage, Aziz, Koper, Michal, Kufel, Joanna, Tollervey, David

During transcription termination by RNA polymerase II on protein-coding genes, the nuclear 5′ exonuclease Rat1/Xrn2 degrades the nascent transcript downstream from the polyadenylation site and...

Roles of the HEAT repeat proteins Utp10 and Utp20 in 40S ribosome maturation (2007)

Dez, Christophe, Dlakic, Mensur, Tollervey, David

A family of HEAT-repeat containing ribosome synthesis factors was previously identified in Saccharomyces cerevisiae. We report the detailed characterization of two of these factors, Utp10 and Utp20,...

Yeast Rrp14p is required for ribosomal subunit synthesis and for correct positioning of the mitotic spindle during mitosis (2007)

Oeffinger, Marlene, Fatica, Alessandro, Rout, Michael P., Tollervey, David

Here we report that Rrp14p/Ykl082p is associated with pre-60S particles and to a lesser extent with earlier 90S pre-ribosomes. Depletion of Rrp14p inhibited pre-rRNA synthesis on both the 40S and 60S...

Nop9 is an RNA binding protein present in pre-40S ribosomes and required for 18S rRNA synthesis in yeast (2007)

Thomson, Emma, Rappsilber, Juri, Tollervey, David

Proteomic analyses in yeast have identified a large number of proteins that are associated with preribosomal particles. However, the product of the yeast ORF YJL010C, herein designated as Nop9,...

Yeast Rrp14p is required for ribosomal subunit synthesis and for correct positioning of the mitotic spindle during mitosis (2007)

Oeffinger, Marlene, Fatica, Alessandro, Rout, Michael P., Tollervey, David

Here we report that Rrp14p/Ykl082p is associated with pre-60S particles and to a lesser extent with earlier 90S pre-ribosomes. Depletion of Rrp14p inhibited pre-rRNA synthesis on both the 40S and 60S...

FLIPing heterokaryons to analyze nucleo-cytoplasmic shuttling of yeast proteins (2006)

Belaya, Katsiaryna, Tollervey, David, Kos, Martin

Nucleo-cytoplasmic shuttling is an important feature of proteins involved in nuclear export/import of RNAs, proteins, and also large ribonucleoprotein complexes such as ribosomes. The vast amount of...

FLIPing heterokaryons to analyze nucleo-cytoplasmic shuttling of yeast proteins (2006)

Belaya, Katsiaryna, Tollervey, David, Kos, Martin

Nucleo-cytoplasmic shuttling is an important feature of proteins involved in nuclear export/import of RNAs, proteins, and also large ribonucleoprotein complexes such as ribosomes. The vast amount of...

FLIPing heterokaryons to analyze nucleo-cytoplasmic shuttling of yeast proteins (2006)

Belaya, Katsiaryna, Tollervey, David, Kos, Martin

Nucleo-cytoplasmic shuttling is an important feature of proteins involved in nuclear export/import of RNAs, proteins, and also large ribonucleoprotein complexes such as ribosomes. The vast amount of...

Nop53p is required for late 60S ribosome subunit maturation and nuclear export in yeast (2005)

THOMSON, EMMA, TOLLERVEY, DAVID

We report that Ypl146cp/Nop53p is associated with pre-60S ribosomal complexes and localized to the nucleolus and nucleoplasm. In cells depleted of Nop53p synthesis of the rRNA components of the 60S...

A pre-ribosome-associated HEAT-repeat protein is required for export of both ribosomal subunits (2004)

Oeffinger, Marlene, Dlakic, Mensur, Tollervey, David

Rrp12p (Ypl012w) is unusual among characterized ribosome synthesis factors in being associated with late precursors to both the 40S and 60S subunits. Rrp12p is predominately nuclear with nucleolar...

PIN domain of Nob1p is required for D-site cleavage in 20S pre-rRNA (2004)

FATICA, ALESSANDRO, TOLLERVEY, DAVID, DLAKIC, MENSUR

Nob1p (Yor056c) is essential for processing of the 20S pre-rRNA to the mature 18S rRNA. It is part of a pre-40S ribosomal particle that is transported to the cytoplasm and subsequently cleaved at the...

A pre-ribosome-associated HEAT-repeat protein is required for export of both ribosomal subunits (2004)

Oeffinger, Marlene, Dlakic, Mensur, Tollervey, David

Rrp12p (Ypl012w) is unusual among characterized ribosome synthesis factors in being associated with late precursors to both the 40S and 60S subunits. Rrp12p is predominately nuclear with nucleolar...

The 'scavenger' m7GpppX pyrophosphatase activity of Dcs1 modulates nutrient-induced responses in yeast (2004)

Malys, Naglis, Carroll, Kathleen, Miyan, Jaleel, Tollervey, David, McCarthy, John E. G.

Dcs1, the m7GpppX pyrophosphatase of Saccharomyces cerevisiae, has been reported to ‘scavenge’ capped 5′ end fragments generated by 3′→5′ mRNA degradation. We now show that the absence of...

PIN domain of Nob1p is required for D-site cleavage in 20S pre-rRNA (2004)

FATICA, ALESSANDRO, TOLLERVEY, DAVID, DLAKIC, MENSUR

Nob1p (Yor056c) is essential for processing of the 20S pre-rRNA to the mature 18S rRNA. It is part of a pre-40S ribosomal particle that is transported to the cytoplasm and subsequently cleaved at the...

A pre-ribosome-associated HEAT-repeat protein is required for export of both ribosomal subunits (2004)

Oeffinger, Marlene, Dlakic, Mensur, Tollervey, David

Rrp12p (Ypl012w) is unusual among characterized ribosome synthesis factors in being associated with late precursors to both the 40S and 60S subunits. Rrp12p is predominately nuclear with nucleolar...

3'-processing of yeast tRNATrp precedes 5'-processing (2003)

KUFEL, JOANNA, TOLLERVEY, DAVID

Previous analyses of eukaryotic pre-tRNAs processing have reported that 5′-cleavage by RNase P precedes 3′-maturation. Here we report that in contrast to all other yeast tRNAs analyzed to date,...

A complex pathway for 3' processing of the yeast U3 snoRNA (2003)

Kufel, Joanna, Allmang, Christine, Verdone, Loredana, Beggs, Jean, Tollervey, David

Mature U3 snoRNA in yeast is generated from the 3′‐extended precursors by endonucleolytic cleavage followed by exonucleolytic trimming. These precursors terminate in poly(U) tracts and are...

The function and synthesis of Ribosomes (2001)

Lafontaine, Denis LJ, Tollervey, David

Structural analyses of the large and small ribosomal subunits have allowed us to think about how they work in more detail than ever before. The mechanisms that underlie ribosomal synthesis,...

The function and synthesis of Ribosomes (2001)

Lafontaine, Denis LJ, Tollervey, David

Structural analyses of the large and small ribosomal subunits have allowed us to think about how they work in more detail than ever before. The mechanisms that underlie ribosomal synthesis,...

Musing on the structural organization of the exosome complex (2000)

Mitchell, Philip, Tollervey, David

The exosome complex of 3′→5′ exoribonucleases functions in both the precise processing of 3′ extended precursor molecules to mature stable RNAs and the complete degradation of other RNAs....

Musing on the structural organization of the exosome complex (2000)

Mitchell, Philip, Tollervey, David

The exosome complex of 3′→5′ exoribonucleases functions in both the precise processing of 3′ extended precursor molecules to mature stable RNAs and the complete degradation of other RNAs....

Degradation of ribosomal RNA precursors by the exosome (2000)

Allmang, Christine, Mitchell, Philip, Petfalski, Elisabeth, Tollervey, David

The yeast exosome is a complex of 3′→5′ exonucleases involved in RNA processing and degradation. All 11 known components of the exosome are required during 3′ end processing of the 5.8S rRNA....

Mutational analysis of an essential binding site for the U3 snoRNA in the 5' external transcribed spacer of yeast pre-rRNA (1994)

Beltrame, Monica, Henry, Yves, Tollervey, David

The small nucleolar RNA U3 is essential for viability in yeast. We have previously shown that U3 can be crosslinked in vivo to the pre-rRNA in the 5' external transcribed spacer (ETS), at +470. This...

Identification and functional analysis of a novel yeast small nucleolar RNA (1993)

Dandekar, Thomas, Tollervey, David

snR31 is a RNA species of 225 nt. which has the trimethyl guanosine cap structure typical of small nuclear RNAs (snRNAs) and yeast small nucleolar RNAs (snoRNAs), and is associated with the nucleolar...

Evolutionary conserved nucleotides within the E.coli 4.5S RNA are required for association with P48 in vitro and for optimal function in vivo (1992)

Wood, Heather, Luirink, Joen, Tollervey, David

E.coli 4.5S RNA is homologous to domain IV of eukaryotic SRP7S RNA, the RNA component of the signal recognition particle. The 4.5S RNA is associated in vivo with a 48kD protein (P48), which is...

Immunoprecipitation distinguishes non-overlapping groups of snRNPs in Schizosaccharomyces pombe (1990)

Tollervey, David, Tessars, Gabriela, Lührmann, Reinhard

The large number of snRNAs in the fission yeast Schizosaccharomyces pombe can be divided into four non-ovetlapping groups by immunoprecipitation with antibodies directed against mammalian snRNP...

Base Pairing between U3 Small Nucleolar RNA and the 5′ End of 18S rRNA Is Required for Pre-rRNA Processing

Sharma, Kishor, Tollervey, David

The loop of a stem structure close to the 5′ end of the 18S rRNA is complementary to the box A region of the U3 small nucleolar RNA (snoRNA). Substitution of the 18S loop nucleotides inhibited...

Genetic and Physical Interactions Involving the Yeast Nuclear Cap-Binding Complex

Fortes, Puri, Kufel, Joanna, Fornerod, Maarten, Polycarpou-Schwarz, Maria, Lafontaine, Denis, Tollervey, David, ...

Yeast strains lacking the yeast nuclear cap-binding complex (yCBC) are viable, although impaired in growth. We have taken advantage of this observation to carry out a genetic screen for components...

Synthesis and Assembly of the Box C+D Small Nucleolar RNPs

Lafontaine, Denis L. J., Tollervey, David

Two core small nucleolar RNP (snoRNP) proteins, Nop1p (fibrillarin in vertebrates) and Nop58p (also known as Nop5p) have previously been reported to be specifically associated with the box C+D class...

Precursors to the U3 Small Nucleolar RNA Lack Small Nucleolar RNP Proteins but Are Stabilized by La Binding

Kufel, Joanna, Allmang, Christine, Chanfreau, Guillaume, Petfalski, Elisabeth, Lafontaine, Denis L. J., Tollervey, David

Almost all small eukaryotic RNAs are processed from transiently stabilized 3′-extended forms. A key question is how and why such intermediates are stabilized and how they can then be processed to...

Dhr1p, a Putative DEAH-Box RNA Helicase, Is Associated with the Box C+D snoRNP U3

Colley, Alan, Beggs, Jean D., Tollervey, David, Lafontaine, Denis L. J.

Putative RNA helicases are involved in most aspects of gene expression. All previously characterized members of the DEAH-box family of putative RNA helicases are involved in pre-mRNA splicing. Here...

Nuclear Export of 60S Ribosomal Subunits Depends on Xpo1p and Requires a Nuclear Export Sequence-Containing Factor, Nmd3p, That Associates with the Large Subunit Protein Rpl10p

Gadal, Olivier, Strauß, Daniela, Kessl, Jacques, Trumpower, Bernard, Tollervey, David, Hurt, Ed

Nuclear export of ribosomes requires a subset of nucleoporins and the Ran system, but specific transport factors have not been identified. Using a large subunit reporter (Rpl25p-eGFP), we have...

Degradation of ribosomal RNA precursors by the exosome

Allmang, Christine, Mitchell, Philip, Petfalski, Elisabeth, Tollervey, David

The yeast exosome is a complex of 3′→5′ exonucleases involved in RNA processing and degradation. All 11 known components of the exosome are required during 3′ end processing of the 5.8S rRNA....

Processing of the Precursors to Small Nucleolar RNAs and rRNAs Requires Common Components

Petfalski, Elisabeth, Dandekar, Thomas, Henry, Yves, Tollervey, David

The genes encoding the small nucleolar RNA (snoRNA) species snR190 and U14 are located close together in the genome of Saccharomyces cerevisiae. Here we report that these two snoRNAs are synthesized...

Yeast 18S rRNA Dimethylase Dim1p: a Quality Control Mechanism in Ribosome Synthesis?

Lafontaine, Denis L. J., Preiss, Thomas, Tollervey, David

One of the few rRNA modifications conserved between bacteria and eukaryotes is the base dimethylation present at the 3′ end of the small subunit rRNA. In the yeast Saccharomyces cerevisiae, this...

Box C/D small nucleolar RNA trafficking involves small nucleolar RNP proteins, nucleolar factors and a novel nuclear domain

Verheggen, Céline, Mouaikel, John, Thiry, Marc, Blanchard, Jean-Marie, Tollervey, David, Bordonné, Remi, ...

Nucleolar localization of box C/D small nucleolar (sno) RNAs requires the box C/D motif and, in vertebrates, involves transit through Cajal bodies (CB). We report that in yeast, overexpression of a...

A nuclear AAA-type ATPase (Rix7p) is required for biogenesis and nuclear export of 60S ribosomal subunits

Gadal, Olivier, Strauß, Daniela, Braspenning, Joris, Hoepfner, Dominic, Petfalski, Elisabeth, Philippsen, Peter, ...

Ribosomal precursor particles are assembled in the nucleolus before export into the cytoplasm. Using a visual assay for nuclear accumulation of 60S subunits, we have isolated several...

60S pre-ribosome formation viewed from assembly in the nucleolus until export to the cytoplasm

Nissan, Tracy A., Baßler, Jochen, Petfalski, Elisabeth, Tollervey, David, Hurt, Ed

60S ribosomes undergo initial assembly in the nucleolus before export to the cytoplasm and recent analyses have identified several nucleolar pre-60S particles. To unravel the steps in the pathway of...

Lsm Proteins Are Required for Normal Processing of Pre-tRNAs and Their Efficient Association with La-Homologous Protein Lhp1p

Kufel, Joanna, Allmang, Christine, Verdone, Loredana, Beggs, Jean D., Tollervey, David

Depletion of any of the five essential proteins Lsm2p to Lsm5p and Lsm8p leads to strong accumulation of all tested unspliced pre-tRNA species, as well as accumulation of 5′ and 3′ unprocessed...

The path from nucleolar 90S to cytoplasmic 40S pre-ribosomes

Schäfer, Thorsten, Strauß, Daniela, Petfalski, Elisabeth, Tollervey, David, Hurt, Ed

Recent reports have increased our knowledge of the consecutive steps during 60S ribosome biogenesis substantially, but 40S subunit formation is less well understood. Here, we investigate the...

Nob1p Is Required for Cleavage of the 3′ End of 18S rRNA

Fatica, Alessandro, Oeffinger, Marlene, Dlakić, Mensur, Tollervey, David

We report the characterization of a novel factor, Nob1p (Yor056c), which is essential for the synthesis of 40S ribosome subunits. Genetic depletion of Nob1p strongly inhibits the processing of the...

Rrp47p Is an Exosome-Associated Protein Required for the 3′ Processing of Stable RNAs

Mitchell, Philip, Petfalski, Elisabeth, Houalla, Rym, Podtelejnikov, Alexandre, Mann, Matthias, Tollervey, David

Related exosome complexes of 3′→5′ exonucleases are present in the nucleus and the cytoplasm. Purification of exosome complexes from whole-cell lysates identified a Mg2+-labile factor present...

Fibrillarin Is Essential for Early Development and Required for Accumulation of an Intron-Encoded Small Nucleolar RNA in the Mouse

Newton, Kathryn, Petfalski, Elisabeth, Tollervey, David, Cáceres, Javier F.

Fibrillarin, a protein component of C/D box small nucleolar ribonucleoproteins (snoRNPs), directs 2′-O-methylation of rRNA and is also involved in other aspects of rRNA processing. A gene trap...

A complex pathway for 3′ processing of the yeast U3 snoRNA

Kufel, Joanna, Allmang, Christine, Verdone, Loredana, Beggs, Jean, Tollervey, David

Mature U3 snoRNA in yeast is generated from the 3′-extended precursors by endonucleolytic cleavage followed by exonucleolytic trimming. These precursors terminate in poly(U) tracts and are normally...

Yeast Nop15p is an RNA-binding protein required for pre-rRNA processing and cytokinesis

Oeffinger, Marlene, Tollervey, David

Nop15p is an essential protein that contains an RNA recognition motif (RRM) and localizes to the nucleoplasm and nucleolus. Cells depleted of Nop15p failed to synthesize the 25S and 5.8S rRNA...

The box H+ACA snoRNAs carry Cbf5p, the putative rRNA pseudouridine synthase

Lafontaine, Denis L.J., Bousquet-Antonelli, Cécile, Henry, Yves, Caizergues-Ferrer, Michèle, Tollervey, David

Many or all of the sites of pseudouridine (Ψ) formation in eukaryotic rRNA are selected by site-specific base-pairing with members of the box H + ACA class of small nucleolar RNAs (snoRNAs)....

The yeast exosome and human PM–Scl are related complexes of 3′ → 5′ exonucleases

Allmang, Christine, Petfalski, Elisabeth, Podtelejnikov, Alexandre, Mann, Matthias, Tollervey, David, Mitchell, Philip

We previously identified a complex of 3′ → 5′ exoribonucleases, designated the exosome, that is expected to play a major role in diverse RNA processing and degradation pathways. Further...

A pre-ribosome-associated HEAT-repeat protein is required for export of both ribosomal subunits

Oeffinger, Marlene, Dlakić, Mensur, Tollervey, David

Rrp12p (Ypl012w) is unusual among characterized ribosome synthesis factors in being associated with late precursors to both the 40S and 60S subunits. Rrp12p is predominately nuclear with nucleolar...

7SL RNA from Schizosaccharomyces pombe is encoded by a single copy essential gene

Ribes, Véronique, Dehoux, Pierre, Tollervey, David

We have identified an abundant ribonucleoprotein particle from Schizosaccharomyces pombe with properties related to those of the vertebrate signal recognition particle (SRP), including cytoplasmic...

The ‘scavenger’ m7GpppX pyrophosphatase activity of Dcs1 modulates nutrient-induced responses in yeast

Malys, Naglis, Carroll, Kathleen, Miyan, Jaleel, Tollervey, David, McCarthy, John E. G.

Dcs1, the m7GpppX pyrophosphatase of Saccharomyces cerevisiae, has been reported to ‘scavenge’ capped 5′ end fragments generated by 3′→5′ mRNA degradation. We now show that the absence of...

Nuclear Pre-mRNA Decapping and 5′ Degradation in Yeast Require the Lsm2-8p Complex

Kufel, Joanna, Bousquet-Antonelli, Cecile, Beggs, Jean D., Tollervey, David

Previous analyses have identified related cytoplasmic Lsm1-7p and nuclear Lsm2-8p complexes. Here we report that mature heat shock and MET mRNAs that are trapped in the nucleus due to a block in mRNA...

Functional link between ribosome formation and biogenesis of iron–sulfur proteins

Yarunin, Alexander, Panse, Vikram Govind, Petfalski, Elisabeth, Dez, Christophe, Tollervey, David, Hurt, Ed

In genetic screens for ribosomal export mutants, we identified CFD1, NBP35 and NAR1 as factors involved in ribosome biogenesis. Notably, these components were recently reported to function in...

A Nuclear Surveillance Pathway for mRNAs with Defective Polyadenylation¶

Milligan, Laura, Torchet, Claire, Allmang, Christine, Shipman, Tracey, Tollervey, David

The pap1-5 mutation in poly(A) polymerase causes rapid depletion of mRNAs at restrictive temperatures. Residual mRNAs are polyadenylated, indicating that Pap1-5p retains at least partial activity. In...

Formation and nuclear export of tRNA, rRNA and mRNA is regulated by the ubiquitin ligase Rsp5p

Neumann, Silvia, Petfalski, Elisabeth, Brügger, Britta, Großhans, Helge, Wieland, Felix, Tollervey, David, ...

The yeast ubiquitin-protein ligase Rsp5p regulates processes as diverse as polII transcription and endocytosis. Here, we identify Rsp5p in a screen for tRNA export (tex) mutants. The tex23-1/rsp5-3...

Yeast Pescadillo is required for multiple activities during 60S ribosomal subunit synthesis.

Oeffinger, Marlene, Leung, Anthony, Lamond, Angus, Tollervey, David, Lueng, Anthony

The Pescadillo protein was identified via a developmental defect and implicated in cell cycle progression. Here we report that human Pescadillo and its yeast homolog (Yph1p or Nop7p) are localized to...

Naf1 p is a box H/ACA snoRNP assembly factor.

Fatica, Alessandro, Dlakić, Mensur, Tollervey, David

Box H/ACA small nucleolar ribonucleoprotein particles (snoRNPs) contain four essential proteins, Cbf5p, Gar1p, Nhp2p, and Nop10p, each of which, with the exception of Gar1p, is required for box H/ACA...

3′-processing of yeast tRNATrp precedes 5′-processing

KUFEL, JOANNA, TOLLERVEY, DAVID

Previous analyses of eukaryotic pre-tRNAs processing have reported that 5′-cleavage by RNase P precedes 3′-maturation. Here we report that in contrast to all other yeast tRNAs analyzed to date,...

Cic1p/Nsa3p is required for synthesis and nuclear export of 60S ribosomal subunits

FATICA, ALESSANDRO, OEFFINGER, MARLENE, TOLLERVEY, DAVID, BOZZONI, IRENE

Cic1p/Nsa3p was previously reported to be associated with the 26S proteasome and required for the degradation of specific substrates, but was also shown to be associated with early pre-60S particles...

PIN domain of Nob1p is required for D-site cleavage in 20S pre-rRNA

FATICA, ALESSANDRO, TOLLERVEY, DAVID, DLAKIĆ, MENSUR

Nob1p (Yor056c) is essential for processing of the 20S pre-rRNA to the mature 18S rRNA. It is part of a pre-40S ribosomal particle that is transported to the cytoplasm and subsequently cleaved at the...

Nop53p is required for late 60S ribosome subunit maturation and nuclear export in yeast

THOMSON, EMMA, TOLLERVEY, DAVID

We report that Ypl146cp/Nop53p is associated with pre-60S ribosomal complexes and localized to the nucleolus and nucleoplasm. In cells depleted of Nop53p synthesis of the rRNA components of the 60S...

The Noc proteins involved in ribosome synthesis and export contain divergent HEAT repeats

DLAKIĆ, MENSUR, TOLLERVEY, DAVID

The Noc1–4p proteins were previously reported to be involved in intranuclear and nucleocytoplasmic transport of pre-ribosomes. Using fold recognition and structural modeling, we show that Noc1–4p...

FLIPing heterokaryons to analyze nucleo-cytoplasmic shuttling of yeast proteins

Belaya, Katsiaryna, Tollervey, David, Koš, Martin

Nucleo-cytoplasmic shuttling is an important feature of proteins involved in nuclear export/import of RNAs, proteins, and also large ribonucleoprotein complexes such as ribosomes. The vast amount of...

Base Pairing between U3 Small Nucleolar RNA and the 5′ End of 18S rRNA Is Required for Pre-rRNA Processing

Sharma, Kishor, Tollervey, David

The loop of a stem structure close to the 5′ end of the 18S rRNA is complementary to the box A region of the U3 small nucleolar RNA (snoRNA). Substitution of the 18S loop nucleotides inhibited...

Genetic and Physical Interactions Involving the Yeast Nuclear Cap-Binding Complex

Fortes, Puri, Kufel, Joanna, Fornerod, Maarten, Polycarpou-Schwarz, Maria, Lafontaine, Denis, Tollervey, David, ...

Yeast strains lacking the yeast nuclear cap-binding complex (yCBC) are viable, although impaired in growth. We have taken advantage of this observation to carry out a genetic screen for components...

Synthesis and Assembly of the Box C+D Small Nucleolar RNPs

Lafontaine, Denis L. J., Tollervey, David

Two core small nucleolar RNP (snoRNP) proteins, Nop1p (fibrillarin in vertebrates) and Nop58p (also known as Nop5p) have previously been reported to be specifically associated with the box C+D class...

Precursors to the U3 Small Nucleolar RNA Lack Small Nucleolar RNP Proteins but Are Stabilized by La Binding

Kufel, Joanna, Allmang, Christine, Chanfreau, Guillaume, Petfalski, Elisabeth, Lafontaine, Denis L. J., Tollervey, David

Almost all small eukaryotic RNAs are processed from transiently stabilized 3′-extended forms. A key question is how and why such intermediates are stabilized and how they can then be processed to...

Dhr1p, a Putative DEAH-Box RNA Helicase, Is Associated with the Box C+D snoRNP U3

Colley, Alan, Beggs, Jean D., Tollervey, David, Lafontaine, Denis L. J.

Putative RNA helicases are involved in most aspects of gene expression. All previously characterized members of the DEAH-box family of putative RNA helicases are involved in pre-mRNA splicing. Here...

Nuclear Export of 60S Ribosomal Subunits Depends on Xpo1p and Requires a Nuclear Export Sequence-Containing Factor, Nmd3p, That Associates with the Large Subunit Protein Rpl10p

Gadal, Olivier, Strauß, Daniela, Kessl, Jacques, Trumpower, Bernard, Tollervey, David, Hurt, Ed

Nuclear export of ribosomes requires a subset of nucleoporins and the Ran system, but specific transport factors have not been identified. Using a large subunit reporter (Rpl25p-eGFP), we have...

Degradation of ribosomal RNA precursors by the exosome

Allmang, Christine, Mitchell, Philip, Petfalski, Elisabeth, Tollervey, David

The yeast exosome is a complex of 3′→5′ exonucleases involved in RNA processing and degradation. All 11 known components of the exosome are required during 3′ end processing of the 5.8S rRNA....

Processing of the Precursors to Small Nucleolar RNAs and rRNAs Requires Common Components

Petfalski, Elisabeth, Dandekar, Thomas, Henry, Yves, Tollervey, David

The genes encoding the small nucleolar RNA (snoRNA) species snR190 and U14 are located close together in the genome of Saccharomyces cerevisiae. Here we report that these two snoRNAs are synthesized...

Yeast 18S rRNA Dimethylase Dim1p: a Quality Control Mechanism in Ribosome Synthesis?

Lafontaine, Denis L. J., Preiss, Thomas, Tollervey, David

One of the few rRNA modifications conserved between bacteria and eukaryotes is the base dimethylation present at the 3′ end of the small subunit rRNA. In the yeast Saccharomyces cerevisiae, this...

Box C/D small nucleolar RNA trafficking involves small nucleolar RNP proteins, nucleolar factors and a novel nuclear domain

Verheggen, Céline, Mouaikel, John, Thiry, Marc, Blanchard, Jean-Marie, Tollervey, David, Bordonné, Remi, ...

Nucleolar localization of box C/D small nucleolar (sno) RNAs requires the box C/D motif and, in vertebrates, involves transit through Cajal bodies (CB). We report that in yeast, overexpression of a...

A nuclear AAA-type ATPase (Rix7p) is required for biogenesis and nuclear export of 60S ribosomal subunits

Gadal, Olivier, Strauß, Daniela, Braspenning, Joris, Hoepfner, Dominic, Petfalski, Elisabeth, Philippsen, Peter, ...

Ribosomal precursor particles are assembled in the nucleolus before export into the cytoplasm. Using a visual assay for nuclear accumulation of 60S subunits, we have isolated several...

60S pre-ribosome formation viewed from assembly in the nucleolus until export to the cytoplasm

Nissan, Tracy A., Baßler, Jochen, Petfalski, Elisabeth, Tollervey, David, Hurt, Ed

60S ribosomes undergo initial assembly in the nucleolus before export to the cytoplasm and recent analyses have identified several nucleolar pre-60S particles. To unravel the steps in the pathway of...

Lsm Proteins Are Required for Normal Processing of Pre-tRNAs and Their Efficient Association with La-Homologous Protein Lhp1p

Kufel, Joanna, Allmang, Christine, Verdone, Loredana, Beggs, Jean D., Tollervey, David

Depletion of any of the five essential proteins Lsm2p to Lsm5p and Lsm8p leads to strong accumulation of all tested unspliced pre-tRNA species, as well as accumulation of 5′ and 3′ unprocessed...

The path from nucleolar 90S to cytoplasmic 40S pre-ribosomes

Schäfer, Thorsten, Strauß, Daniela, Petfalski, Elisabeth, Tollervey, David, Hurt, Ed

Recent reports have increased our knowledge of the consecutive steps during 60S ribosome biogenesis substantially, but 40S subunit formation is less well understood. Here, we investigate the...

Nob1p Is Required for Cleavage of the 3′ End of 18S rRNA

Fatica, Alessandro, Oeffinger, Marlene, Dlakić, Mensur, Tollervey, David

We report the characterization of a novel factor, Nob1p (Yor056c), which is essential for the synthesis of 40S ribosome subunits. Genetic depletion of Nob1p strongly inhibits the processing of the...

Rrp47p Is an Exosome-Associated Protein Required for the 3′ Processing of Stable RNAs

Mitchell, Philip, Petfalski, Elisabeth, Houalla, Rym, Podtelejnikov, Alexandre, Mann, Matthias, Tollervey, David

Related exosome complexes of 3′→5′ exonucleases are present in the nucleus and the cytoplasm. Purification of exosome complexes from whole-cell lysates identified a Mg2+-labile factor present...

Fibrillarin Is Essential for Early Development and Required for Accumulation of an Intron-Encoded Small Nucleolar RNA in the Mouse

Newton, Kathryn, Petfalski, Elisabeth, Tollervey, David, Cáceres, Javier F.

Fibrillarin, a protein component of C/D box small nucleolar ribonucleoproteins (snoRNPs), directs 2′-O-methylation of rRNA and is also involved in other aspects of rRNA processing. A gene trap...

A complex pathway for 3′ processing of the yeast U3 snoRNA

Kufel, Joanna, Allmang, Christine, Verdone, Loredana, Beggs, Jean, Tollervey, David

Mature U3 snoRNA in yeast is generated from the 3′-extended precursors by endonucleolytic cleavage followed by exonucleolytic trimming. These precursors terminate in poly(U) tracts and are normally...

Yeast Nop15p is an RNA-binding protein required for pre-rRNA processing and cytokinesis

Oeffinger, Marlene, Tollervey, David

Nop15p is an essential protein that contains an RNA recognition motif (RRM) and localizes to the nucleoplasm and nucleolus. Cells depleted of Nop15p failed to synthesize the 25S and 5.8S rRNA...

The box H+ACA snoRNAs carry Cbf5p, the putative rRNA pseudouridine synthase

Lafontaine, Denis L.J., Bousquet-Antonelli, Cécile, Henry, Yves, Caizergues-Ferrer, Michèle, Tollervey, David

Many or all of the sites of pseudouridine (Ψ) formation in eukaryotic rRNA are selected by site-specific base-pairing with members of the box H + ACA class of small nucleolar RNAs (snoRNAs)....

The yeast exosome and human PM–Scl are related complexes of 3′ → 5′ exonucleases

Allmang, Christine, Petfalski, Elisabeth, Podtelejnikov, Alexandre, Mann, Matthias, Tollervey, David, Mitchell, Philip

We previously identified a complex of 3′ → 5′ exoribonucleases, designated the exosome, that is expected to play a major role in diverse RNA processing and degradation pathways. Further...

A pre-ribosome-associated HEAT-repeat protein is required for export of both ribosomal subunits

Oeffinger, Marlene, Dlakić, Mensur, Tollervey, David

Rrp12p (Ypl012w) is unusual among characterized ribosome synthesis factors in being associated with late precursors to both the 40S and 60S subunits. Rrp12p is predominately nuclear with nucleolar...

7SL RNA from Schizosaccharomyces pombe is encoded by a single copy essential gene

Ribes, Véronique, Dehoux, Pierre, Tollervey, David

We have identified an abundant ribonucleoprotein particle from Schizosaccharomyces pombe with properties related to those of the vertebrate signal recognition particle (SRP), including cytoplasmic...

The ‘scavenger’ m7GpppX pyrophosphatase activity of Dcs1 modulates nutrient-induced responses in yeast

Malys, Naglis, Carroll, Kathleen, Miyan, Jaleel, Tollervey, David, McCarthy, John E. G.

Dcs1, the m7GpppX pyrophosphatase of Saccharomyces cerevisiae, has been reported to ‘scavenge’ capped 5′ end fragments generated by 3′→5′ mRNA degradation. We now show that the absence of...

Nuclear Pre-mRNA Decapping and 5′ Degradation in Yeast Require the Lsm2-8p Complex

Kufel, Joanna, Bousquet-Antonelli, Cecile, Beggs, Jean D., Tollervey, David

Previous analyses have identified related cytoplasmic Lsm1-7p and nuclear Lsm2-8p complexes. Here we report that mature heat shock and MET mRNAs that are trapped in the nucleus due to a block in mRNA...

Functional link between ribosome formation and biogenesis of iron–sulfur proteins

Yarunin, Alexander, Panse, Vikram Govind, Petfalski, Elisabeth, Dez, Christophe, Tollervey, David, Hurt, Ed

In genetic screens for ribosomal export mutants, we identified CFD1, NBP35 and NAR1 as factors involved in ribosome biogenesis. Notably, these components were recently reported to function in...

A Nuclear Surveillance Pathway for mRNAs with Defective Polyadenylation¶

Milligan, Laura, Torchet, Claire, Allmang, Christine, Shipman, Tracey, Tollervey, David

The pap1-5 mutation in poly(A) polymerase causes rapid depletion of mRNAs at restrictive temperatures. Residual mRNAs are polyadenylated, indicating that Pap1-5p retains at least partial activity. In...

Formation and nuclear export of tRNA, rRNA and mRNA is regulated by the ubiquitin ligase Rsp5p

Neumann, Silvia, Petfalski, Elisabeth, Brügger, Britta, Großhans, Helge, Wieland, Felix, Tollervey, David, ...

The yeast ubiquitin-protein ligase Rsp5p regulates processes as diverse as polII transcription and endocytosis. Here, we identify Rsp5p in a screen for tRNA export (tex) mutants. The tex23-1/rsp5-3...

Yeast Pescadillo is required for multiple activities during 60S ribosomal subunit synthesis.

Oeffinger, Marlene, Leung, Anthony, Lamond, Angus, Tollervey, David, Lueng, Anthony

The Pescadillo protein was identified via a developmental defect and implicated in cell cycle progression. Here we report that human Pescadillo and its yeast homolog (Yph1p or Nop7p) are localized to...

Naf1 p is a box H/ACA snoRNP assembly factor.

Fatica, Alessandro, Dlakić, Mensur, Tollervey, David

Box H/ACA small nucleolar ribonucleoprotein particles (snoRNPs) contain four essential proteins, Cbf5p, Gar1p, Nhp2p, and Nop10p, each of which, with the exception of Gar1p, is required for box H/ACA...

3′-processing of yeast tRNATrp precedes 5′-processing

KUFEL, JOANNA, TOLLERVEY, DAVID

Previous analyses of eukaryotic pre-tRNAs processing have reported that 5′-cleavage by RNase P precedes 3′-maturation. Here we report that in contrast to all other yeast tRNAs analyzed to date,...

Cic1p/Nsa3p is required for synthesis and nuclear export of 60S ribosomal subunits

FATICA, ALESSANDRO, OEFFINGER, MARLENE, TOLLERVEY, DAVID, BOZZONI, IRENE

Cic1p/Nsa3p was previously reported to be associated with the 26S proteasome and required for the degradation of specific substrates, but was also shown to be associated with early pre-60S particles...

PIN domain of Nob1p is required for D-site cleavage in 20S pre-rRNA

FATICA, ALESSANDRO, TOLLERVEY, DAVID, DLAKIĆ, MENSUR

Nob1p (Yor056c) is essential for processing of the 20S pre-rRNA to the mature 18S rRNA. It is part of a pre-40S ribosomal particle that is transported to the cytoplasm and subsequently cleaved at the...

Nop53p is required for late 60S ribosome subunit maturation and nuclear export in yeast

THOMSON, EMMA, TOLLERVEY, DAVID

We report that Ypl146cp/Nop53p is associated with pre-60S ribosomal complexes and localized to the nucleolus and nucleoplasm. In cells depleted of Nop53p synthesis of the rRNA components of the 60S...

The Noc proteins involved in ribosome synthesis and export contain divergent HEAT repeats

DLAKIĆ, MENSUR, TOLLERVEY, DAVID

The Noc1–4p proteins were previously reported to be involved in intranuclear and nucleocytoplasmic transport of pre-ribosomes. Using fold recognition and structural modeling, we show that Noc1–4p...

FLIPing heterokaryons to analyze nucleo-cytoplasmic shuttling of yeast proteins

Belaya, Katsiaryna, Tollervey, David, Koš, Martin

Nucleo-cytoplasmic shuttling is an important feature of proteins involved in nuclear export/import of RNAs, proteins, and also large ribonucleoprotein complexes such as ribosomes. The vast amount of...

Yeast Trf5p is a nuclear poly(A) polymerase

Houseley, Jonathan, Tollervey, David

Recent analyses have shown that the activity of the yeast nuclear exosome is stimulated by the Trf4p–Air1/2p–Mtr4p polyadenylation (TRAMP) complex. Here, we report that strains lacking the Rrp6p...

Surveillance of nuclear-restricted pre-ribosomes within a subnucleolar region of Saccharomyces cerevisiae

Dez, Christophe, Houseley, Jonathan, Tollervey, David

We previously hypothesized that HEAT-repeat (Huntington, elongation A subunit, TOR) ribosome synthesis factors function in ribosome export. We report that the HEAT-repeat protein Sda1p is a component...

Yeast Rrp14p is required for ribosomal subunit synthesis and for correct positioning of the mitotic spindle during mitosis

Oeffinger, Marlene, Fatica, Alessandro, Rout, Michael P., Tollervey, David

Here we report that Rrp14p/Ykl082p is associated with pre-60S particles and to a lesser extent with earlier 90S pre-ribosomes. Depletion of Rrp14p inhibited pre-rRNA synthesis on both the 40S and 60S...

Depletion of the Yeast Nuclear Exosome Subunit Rrp6 Results in Accumulation of Polyadenylated RNAs in a Discrete Domain within the Nucleolus▿

Carneiro, Tiago, Carvalho, Célia, Braga, José, Rino, José, Milligan, Laura, Tollervey, David, ...

Recent data reveal that a substantial fraction of transcripts generated by RNA polymerases I, II, and III are rapidly degraded in the nucleus by the combined action of the exosome and a noncanonical...

Roles of the HEAT repeat proteins Utp10 and Utp20 in 40S ribosome maturation

Dez, Christophe, Dlakić, Mensur, Tollervey, David

A family of HEAT-repeat containing ribosome synthesis factors was previously identified in Saccharomyces cerevisiae. We report the detailed characterization of two of these factors, Utp10 and Utp20,...

The yeast ribosome synthesis factor Emg1 is a novel member of the superfamily of alpha/beta knot fold methyltransferases

Leulliot, Nicolas, Bohnsack, Markus T., Graille, Marc, Tollervey, David, Van Tilbeurgh, Herman

Emg1 was previously shown to be required for maturation of the 18S rRNA and biogenesis of the 40S ribosomal subunit. Here we report the determination of the crystal structure of Emg1 at 2 Å...

A Novel In Vivo Assay Reveals Inhibition of Ribosomal Nuclear Export in Ran-Cycle and Nucleoporin Mutants

Hurt, Ed, Hannus, Stefan, Schmelzl, Birgit, Lau, Denise, Tollervey, David, Simos, George

To identify components involved in the nuclear export of ribosomes in yeast, we developed an in vivo assay exploiting a green fluorescent protein (GFP)-tagged version of ribosomal protein L25. After...

Rlp7p is associated with 60S preribosomes, restricted to the granular component of the nucleolus, and required for pre-rRNA processing

Gadal, Olivier, Strauss, Daniela, Petfalski, Elisabeth, Gleizes, Pierre-Emmanuel, Gas, Nicole, Tollervey, David, ...

Many analyses have examined subnucleolar structures in eukaryotic cells, but the relationship between morphological structures, pre-rRNA processing, and ribosomal particle assembly has remained...

The mRNA encoding the yeast ARE-binding protein Cth2 is generated by a novel 3′ processing pathway

Ciais, Delphine, Bohnsack, Markus T., Tollervey, David

Microarray analyses of mRNAs over-expressed in strains lacking the nuclear exosome component Rrp6 identified the transcript encoding the ARE-binding protein Cth2, which functions in cytoplasmic mRNA...

Nop9 is an RNA binding protein present in pre-40S ribosomes and required for 18S rRNA synthesis in yeast

Thomson, Emma, Rappsilber, Juri, Tollervey, David

Proteomic analyses in yeast have identified a large number of proteins that are associated with preribosomal particles. However, the product of the yeast ORF YJL010C, herein designated as Nop9,...

Inactivation of Cleavage Factor I Components Rna14p and Rna15p Induces Sequestration of Small Nucleolar Ribonucleoproteins at Discrete Sites in the Nucleus

Carneiro, Tiago, Carvalho, Célia, Braga, José, Rino, José, Milligan, Laura, Tollervey, David, ...

Small nucleolar RNAs (snoRNAs) associate with specific proteins forming small nucleolar ribonucleoprotein (snoRNP) particles, which are essential for ribosome biogenesis. The snoRNAs are transcribed,...

Efficient termination of transcription by RNA polymerase I requires the 5′ exonuclease Rat1 in yeast

El Hage, Aziz, Koper, Michal, Kufel, Joanna, Tollervey, David

During transcription termination by RNA polymerase II on protein-coding genes, the nuclear 5′ exonuclease Rat1/Xrn2 degrades the nascent transcript downstream from the polyadenylation site and...

Trf4 targets ncRNAs from telomeric and rDNA spacer regions and functions in rDNA copy number control

Houseley, Jonathan, Kotovic, Kimberly, El Hage, Aziz, Tollervey, David

Trf4 is the poly(A) polymerase component of TRAMP4, which stimulates nuclear RNA degradation by the exosome. We report that in Saccharomyces cerevisiae strains lacking Trf4, cryptic transcripts are...

An Endoribonuclease Functionally Linked to Perinuclear mRNP Quality Control Associates with the Nuclear Pore Complexes

Skružný, Michal, Schneider, Claudia, Rácz, Attila, Weng, Julan, Tollervey, David, Hurt, Ed

Nuclear mRNA export is a crucial step in eukaryotic gene expression, which is in yeast coupled to cotranscriptional messenger ribonucleoprotein particle (mRNP) assembly and surveillance. Several...

A Yeast Exosome Cofactor, Mpp6, Functions in RNA Surveillance and in the Degradation of Noncoding RNA Transcripts▿

Milligan, Laura, Decourty, Laurence, Saveanu, Cosmin, Rappsilber, Juri, Ceulemans, Hugo, Jacquier, Alain, ...

A genome-wide screen for synthetic lethal (SL) interactions with loss of the nuclear exosome cofactors Rrp47/Lrp1 or Air1 identified 3′→5′ exonucleases, the THO complex required for mRNP...

The N-terminal PIN domain of the exosome subunit Rrp44 harbors endonuclease activity and tethers Rrp44 to the yeast core exosome

Schneider, Claudia, Leung, Eileen, Brown, Jeremy, Tollervey, David

Nuclear and cytoplasmic forms of the yeast exosome share 10 components, of which only Rrp44/Dis3 is believed to possess 3′ exonuclease activity. We report that expression only of Rrp44 lacking...

Quantitative analysis of snoRNA association with pre-ribosomes and release of snR30 by Rok1 helicase

Bohnsack, Markus T, Kos, Martin, Tollervey, David

In yeast, three small nucleolar RNAs (snoRNAs) are essential for the processing of pre-ribosomal RNA—U3, U14 and snR30—whereas 72 non-essential snoRNAs direct site-specific modification of...

Identification of protein binding sites on U3 snoRNA and pre-rRNA by UV cross-linking and high-throughput analysis of cDNAs

Granneman, Sander, Kudla, Grzegorz, Petfalski, Elisabeth, Tollervey, David

The U3 small nucleolar ribonucleoprotein (snoRNP) plays an essential role in ribosome biogenesis but, like many RNA–protein complexes, its architecture is poorly understood. To address this...