The use of simple model systems to study spliceosomal catalysis. (2009)
Valadkhan, Saba, Manley, James L.
Since direct analysis of many aspects of spliceosomal function is greatly hindered by the daunting complexity of the spliceosome, the development of functionally validated simple model systems can be...
Transcription termination by nuclear RNA polymerases (2009)
Richard, Patricia, Manley, James L.
Gene transcription in the cell nucleus is a complex and highly regulated process. Transcription in eukaryotes requires three distinct RNA polymerases, each of which employs its own mechanisms for...
Beckerman, Rachel, Donner, Aaron J., Mattia, Melissa, Peart, Melissa J., Manley, James L., Espinosa, Joaquin M., ...
We reported previously that when cells are arrested in S phase, a subset of p53 target genes fails to be strongly induced despite the presence of high levels of p53. When DNA replication is...
The TET Family of Proteins: Functions and Roles in Disease (2009)
Tan, Adelene Y., Manley, James L.
Translocated in liposarcoma, Ewing's sarcoma and TATA-binding protein-associated factor 15 constitute an interesting and important family of proteins known as the TET proteins. The proteins function...
The 3' processing factor CstF functions in the DNA repair response (2008)
Mirkin, Nurit, Fonseca, Danae, Mohammed, Samia, Cevher, Murat A., Manley, James L., Kleiman, Frida E.
Following DNA damage, mRNA levels decrease, reflecting a coordinated interaction of the DNA repair, transcription and RNA processing machineries. In this study, we provide evidence that transcription...
Sumoylation regulates multiple aspects of mammalian poly(A) polymerase function (2008)
Vethantham, Vasupradha, Rao, Nishta, Manley, James L.
The addition of the poly(A) tail to the ends of eukaryotic mRNAs is catalyzed by poly(A) polymerase (PAP). PAP activity is known to be highly regulated, for example, by alternative splicing and...
Concurrent splicing and transcription are not sufficient to enhance splicing efficiency (2007)
Lazarev, Denis, Manley, James L.
The concept of a tight integration of transcription and splicing of mRNA precursors has been supported with increasing evidence in recent years. However, the mechanism and functional consequences of...
Valadkhan, Saba, Mohammadi, Afshin, Wachtel, Chaim, Manley, James L.
Splicing of introns from mRNA precursors is a two-step reaction performed by the spliceosome, an immense cellular machine consisting of over 200 different proteins and five small RNAs (snRNAs). We...
Pin1 modulates RNA polymerase II activity during the transcription cycle (2007)
The C-terminal domain of the RNA polymerase (RNAP) II largest subunit (CTD) plays a critical role in coordinating multiple events in pre-mRNA transcription and processing. Previously we reported that...
The RNA binding protein RNPS1 alleviates ASF/SF2 depletion-induced genomic instability (2007)
Li, Xialu, Niu, Tianhui, Manley, James L.
Formation of transcription-induced R-loops poses a critical threat to genomic integrity throughout evolution. We have recently shown that the SR protein ASF/SF2 prevents R-loop formation in...
Valadkhan, Saba, Mohammadi, Afshin, Wachtel, Chaim, Manley, James L.
Splicing of introns from mRNA precursors is a two-step reaction performed by the spliceosome, an immense cellular machine consisting of over 200 different proteins and five small RNAs (snRNAs). We...
hnRNP A1 functions with specificity in repression of SMN2 exon 7 splicing (2007)
Kashima, Tsuyoshi, Rao, Nishta, David, Charles J., Manley, James L.
Homozygous deletion or mutation of the survival of motor neuron 1 gene (SMN1) causes spinal muscular atrophy. SMN1 has been duplicated in humans to create SMN2, which produces a low level of...
Cotranscriptional processes and their influence on genome stability (2006)
Numerous studies support the idea that the complex process of gene expression is composed of multiple highly coordinated and integrated steps. While such an extensive coupling ensures the efficiency...
Multiple Properties of the Splicing Repressor SRp38 Distinguish It from Typical SR Proteins (2005)
Chanseok Shin, Frida E. Kleiman, James L. Manley
The SR protein SRp38 is a general splicing repressor that is activated by dephosphorylation during mitosis and in response to heat shock. Here we describe experiments that provide insights into the...
Li, Xialu, Wang, Jin, Manley, James L.
ASF/SF2 is an SR protein splicing factor that participates in constitutive and alternative pre-mRNA splicing and is essential for cell viability. Using a genetically modified chicken B-cell line,...
Li, Xialu, Wang, Jin, Manley, James L.
ASF/SF2 is an SR protein splicing factor that participates in constitutive and alternative pre-mRNA splicing and is essential for cell viability. Using a genetically modified chicken B-cell line,...
Kleiman, Frida E., Wu-Baer, Foon, Fonseca, Danae, Kaneko, Syuzo, Baer, Richard, Manley, James L.
Mammalian cells exhibit a complex response to DNA damage. The tumor suppressor BRCA1 and associated protein BARD1 are thought to play an important role in this response, and our previous work...
Li, Xialu, Wang, Jin, Manley, James L.
ASF/SF2 is an SR protein splicing factor that participates in constitutive and alternative pre-mRNA splicing and is essential for cell viability. Using a genetically modified chicken B-cell line,...
Evidence that polyadenylation factor CPSF-73 is the mRNA 3' processing endonuclease (2004)
RYAN, KEVIN, CALVO, OLGA, MANLEY, JAMES L.
Generation of the polyadenylated 3′ end of an mRNA requires an endonucleolytic cleavage followed by synthesis of the poly(A) tail. Despite the seeming simplicity of the reaction, more than a dozen...
Characterization of the catalytic activity of U2 and U6 snRNAs (2003)
VALADKHAN, SABA, MANLEY, JAMES L.
Removal of introns from pre-messenger RNAs in eukaryotes is carried out by the spliceosome, an assembly of a large number of proteins and five small nuclear RNAs (snRNAs). We showed previously that...
Pin1 modulates the structure and function of human RNA polymerase II (2003)
Xu, Yu-Xin, Hirose, Yutaka, Zhou, Xiao Zhen, Lu, Kun Ping, Manley, James L.
The C-terminal domain of the RNA polymerase (RNAP) II largest subunit (CTD) plays critical roles both in transcription of mRNA precursors and in the processing reactions needed to form mature mRNAs....
Pin1 modulates the structure and function of human RNA polymerase II (2003)
Xu, Yu-Xin, Hirose, Yutaka, Zhou, Xiao Zhen, Lu, Kun Ping, Manley, James L.
The C-terminal domain of the RNA polymerase (RNAP) II largest subunit (CTD) plays critical roles both in transcription of mRNA precursors and in the processing reactions needed to form mature mRNAs....
Lemaire, Raphael, Prasad, Jayendra, Kashima, Tsuyoshi, Gustafson, Jennifer, Manley, James L., Lafyatis, Robert
Hatton, Lee S., Eloranta, Jyrki J., Figueiredo, Luisa M., Takagaki, Yoshio, Manley, James L., OHare, Kevin
During mRNA 3′ end formation, cleavage stimulation factor (CstF) binds to a GU-rich sequence downstream from the polyadenylation site and helps to stabilise the binding of cleavage-polyadenylation...
High mobility group protein-1 (HMG-1) is a unique activator of p53 (1998)
Jayaraman, Lata, Moorthy, Narayani Chandra, Murthy, Kanneganti G.K., Manley, James L., Bustin, Michael, Prives, Carol
Moreira, Alexandra, Takagaki, Yoshio, Brackenridge, Simon, Wollerton, Matthew, Manley, James L., Proudfoot, Nicholas J.
Jamison, Sharon F., Pasman, Zvi, Wang, Jin, Will, Cindy, Lührmann, Reinhard, Manley, James L., ...
Members of the SR family of proteins, can collaborate with U1 snRNP In the recognition of 5′ splice sites in pre-messenger RNAs. We have previously shown that purified U1 snRNP and ASF/SF2 form a...
Woppmann, Andreas, Will, Cindy L., Kornstädt, Ute, Zuo, Ping, Manley, James L., Lührmann, Reinhard
The U1 snRNP-specific 70K protein is one of the few snRNP proteins from higher eukaryotic cells that is phosphorylated In vivo (1,2). Immunoaffinity purified spliceosomal snRNPs (U1, U2, U5, and...
Sequence specificity of the human mRNA N6-adenosine methylase in vitro (1990)
Harper, Joan E., Miceli, Sheila M., Roberts, Richard J., Manley, James L.
N6-adenosine methylation is a frequent modification of mRNAs and their precursors, but little is known about the mechanism of the reaction or the function of the modification. To explore these...
Pan, Zhen-Qiang, Ge, Hui, Fu, Xin-Yuan, Manley, James L., Prives, Carol
We have investigated the roles of U1 and U2 snRNP particles in SV40 pre-mRNA splicing by oligonucleotide-targeted degradation of U1 or U2 snRNAs in Xenopus laevis oocytes. Microinjection of...
RNA polymerase II terminates transcription in vitro in the SV40 origin region (1987)
Grass, David S., Jove, Richard, Manley, James L.
To begin to study signals on DNA that can cause mammalian RNA polymerase II to terminate transcription, we examined the RNA transcripts produced from a number of different DNA templates in a HeLa...
In vitro splicing of simian virus 40 early pre mRNA (1986)
Oble, Jonathan C.S., Rives, Carol, Manley, James L.
The products of splicing of simian virus 40 early pre mRNA in HeLa cell nuclear extracts have been characterized. Of the two alternative splicing patterns exhibited by this precursor in vivo, which...
Transcription of methylated eukaryotic viral genes in a soluble in vitro system (1984)
Jove, Richard, Sperber, David E., Manley, James L.
SV40 and adenovirus-2 (Ad2) recombinant plasmids containing long segments of poly(dC-dG) cloned adjacent to transcription control regions were methylated in vitro with Hhal methylase and transcribed...
The function of guanosine residues surrounding the TATA box of the adenovirus 2 major late promoter (MLP) in promoting efficient transcription initiation and in selecting a specific transcription...
Thesis (M.S.E.H.)--East Tennessee State University, 1974.
Koffa, Maria D., Graham, Sheila V., Takagaki, Yoshio, Manley, James L., Clements, J. Barklie
In human papillomaviruses, expression of the late genes L1 and L2, encoding the capsid proteins, is restricted to the upper layers of the infected epithelium. A 79-nt GU-rich negative regulatory...
Shen, Baohe, Liu, H., Skolnik, Edward Y., Manley, James L.
Signaling through the Toll receptor is required for dorsal/ventral polarity in Drosophila embryos, and also plays an evolutionarily conserved role in the immune response. Upon ligand binding, Toll...
Prasad, Jayendra, Colwill, Karen, Pawson, Tony, Manley, James L.
The splicing of mammalian mRNA precursors requires both protein phosphorylation and dephosphorylation, likely involving modification of members of the SR protein family of splicing factors. Several...
Complex Protein Interactions within the Human Polyadenylation Machinery Identify a Novel Component
Takagaki, Yoshio, Manley, James L.
Polyadenylation of mRNA precursors is a two-step reaction requiring multiple protein factors. Cleavage stimulation factor (CstF) is a heterotrimer necessary for the first step, endonucleolytic...
We have employed gene targeting coupled with conditional expression to construct a chicken DT40 cell line in which a tetracycline (Tet)-repressible promoter is exclusively responsible for expression...
Poly(A) Polymerase Phosphorylation Is Dependent on Novel Interactions with Cyclins
Bond, Gareth L., Prives, Carol, Manley, James L.
We have previously shown that poly(A) polymerase (PAP) is negatively regulated by cyclin B-cdc2 kinase hyperphosphorylation in the M phase of the cell cycle. Here we show that cyclin B1 binds PAP...
Um, Moonkyoung, Yamauchi, Jun, Kato, Shigeaki, Manley, James L.
TATA-binding protein (TBP) is a key general transcription factor required for transcription by all three nuclear RNA polymerases. Although it has been intensively analyzed in vitro and in...
Topalian, Suzanne L., Kaneko, Syuzo, Gonzales, Monica I., Bond, Gareth L., Ward, Yvona, Manley, James L.
Poly(A) polymerase (PAP) plays an essential role in polyadenylation of mRNA precursors, and it has long been thought that mammalian cells contain only a single PAP gene. We describe here the...
Hsp27 Enhances Recovery of Splicing as well as Rephosphorylation of SRp38 after Heat Shock
Marin-Vinader, Laura, Shin, Chanseok, Onnekink, Carla, Manley, James L., Lubsen, Nicolette H.
A heat stress causes a rapid inhibition of splicing. Exogenous expression of Hsp27 did not prevent that inhibition but enhanced the recovery of splicing afterward. Another small heat shock protein,...
Hatton, Lee S., Eloranta, Jyrki J., Figueiredo, Luisa M., Takagaki, Yoshio, Manley, James L., O’Hare, Kevin
During mRNA 3′ end formation, cleavage stimulation factor (CstF) binds to a GU-rich sequence downstream from the polyadenylation site and helps to stabilise the binding of cleavage-polyadenylation...
The Drosophila homeodomain protein Even-skipped (Eve) is a transcriptional repressor, and previous studies have suggested that it functions by interfering with the basal transcription machinery. Here...
Deregulation of Poly(A) Polymerase Interferes with Cell Growth
Zhao, Wenqing, Manley, James L.
Vertebrate poly(A) polymerase (PAP) contains a catalytic domain and a C-terminal Ser-Thr-rich regulatory region. Consensus and nonconsensus cyclin-dependent kinase (cdk) sites are conserved in the...
Requirements of the RNA Polymerase II C-Terminal Domain for Reconstituting Pre-mRNA 3′ Cleavage
Ryan, Kevin, Murthy, Kanneganti G. K., Kaneko, Syuzo, Manley, James L.
RNA polymerase II (RNAP II) has previously been shown to be required for the pre-mRNA polyadenylation cleavage reaction in vitro. This activity was found to reside solely in the C-terminal domain...
ASAP, a Novel Protein Complex Involved in RNA Processing and Apoptosis
Schwerk, Christian, Prasad, Jayendra, Degenhardt, Kurt, Erdjument-Bromage, Hediye, White, Eileen, Tempst, Paul, ...
Different isoforms of a protein complex termed the apoptosis- and splicing-associated protein (ASAP) were isolated from HeLa cell extract. ASAP complexes are composed of the polypeptides SAP18 and...
Regulation and Substrate Specificity of the SR Protein Kinase Clk/Sty
Prasad, Jayendra, Manley, James L.
SR proteins constitute a family of splicing factors that play key roles in both constitutive and regulated splicing in metazoan organisms. The proteins are extensively phosphorylated, and kinases...
Gene-specific transcriptional activation is a multistep process that requires numerous protein factors and DNA elements, including enhancers and the core promoter. To investigate the roles of core...
Pin1 modulates the structure and function of human RNA polymerase II
Xu, Yu-Xin, Hirose, Yutaka, Zhou, Xiao Zhen, Lu, Kun Ping, Manley, James L.
The C-terminal domain of the RNA polymerase (RNAP) II largest subunit (CTD) plays critical roles both in transcription of mRNA precursors and in the processing reactions needed to form mature mRNAs....
High mobility group protein-1 (HMG-1) is a unique activator of p53
Jayaraman, Lata, Moorthy, Narayani Chandra, Murthy, Kanneganti G.K., Manley, James L., Bustin, Michael, Prives, Carol
The binding of p53 protein to DNA is stimulated by its interaction with covalent as well as noncovalent modifiers. We report the identification of a factor from HeLa nuclear extracts that activates...
Phosphorylated RNA polymerase II stimulates pre-mRNA splicing
Hirose, Yutaka, Tacke, Roland, Manley, James L.
RNA polymerase II (RNAP II) is responsible for transcription of mRNA precursors in eukaryotic cells. Recent studies, however, have suggested that RNAP II also participates in subsequent RNA...
Wang, Jin, Xiao, Shou-Hua, Manley, James L.
The SR proteins constitute a family of splicing factors, highly conserved in metazoans, that contain one or two amino-terminal RNA-binding domains (RBDs) and a region enriched in arginine/serine...
Moreira, Alexandra, Takagaki, Yoshio, Brackenridge, Simon, Wollerton, Matthew, Manley, James L., Proudfoot, Nicholas J.
The poly(A) signal of the C2 complement gene is unusual in that it possesses an upstream sequence element (USE) required for full activity in vivo. We describe here in vitro experiments demonstrating...
Millhouse, Scott, Manley, James L.
RNA polymerase II, and specifically the C-terminal domain (CTD) of its largest subunit, has been demonstrated to play important roles in capping, splicing, and 3′ processing of mRNA precursors. But...
The transcriptional coactivator PC4/Sub1 has multiple functions in RNA polymerase II transcription
Transcription and processing of mRNA precursors are coordinated events that require numerous complex interactions to ensure that they are successfully executed. We described previously an unexpected...
BRCA1/BARD1 inhibition of mRNA 3′ processing involves targeted degradation of RNA polymerase II
Kleiman, Frida E., Wu-Baer, Foon, Fonseca, Danae, Kaneko, Syuzo, Baer, Richard, Manley, James L.
Mammalian cells exhibit a complex response to DNA damage. The tumor suppressor BRCA1 and associated protein BARD1 are thought to play an important role in this response, and our previous work...
Multiple Properties of the Splicing Repressor SRp38 Distinguish It from Typical SR Proteins
Shin, Chanseok, Kleiman, Frida E., Manley, James L.
The SR protein SRp38 is a general splicing repressor that is activated by dephosphorylation during mitosis and in response to heat shock. Here we describe experiments that provide insights into the...
Characterization of the catalytic activity of U2 and U6 snRNAs
VALADKHAN, SABA, MANLEY, JAMES L.
Removal of introns from pre-messenger RNAs in eukaryotes is carried out by the spliceosome, an assembly of a large number of proteins and five small nuclear RNAs (snRNAs). We showed previously that...
Evidence that polyadenylation factor CPSF-73 is the mRNA 3′ processing endonuclease
RYAN, KEVIN, CALVO, OLGA, MANLEY, JAMES L.
Generation of the polyadenylated 3′ end of an mRNA requires an endonucleolytic cleavage followed by synthesis of the poly(A) tail. Despite the seeming simplicity of the reaction, more than a dozen...
Lemaire, Raphael, Prasad, Jayendra, Kashima, Tsuyoshi, Gustafson, Jennifer, Manley, James L., Lafyatis, Robert
Pre-mRNA splicing is a widely used regulatory mechanism for controlling gene expression, and a family of conserved proteins, SR proteins, participate in both constitutive and alternative splicing....
Li, Xialu, Wang, Jin, Manley, James L.
ASF/SF2 is an SR protein splicing factor that participates in constitutive and alternative pre-mRNA splicing and is essential for cell viability. Using a genetically modified chicken B-cell line,...
Koffa, Maria D., Graham, Sheila V., Takagaki, Yoshio, Manley, James L., Clements, J. Barklie
In human papillomaviruses, expression of the late genes L1 and L2, encoding the capsid proteins, is restricted to the upper layers of the infected epithelium. A 79-nt GU-rich negative regulatory...
Shen, Baohe, Liu, H., Skolnik, Edward Y., Manley, James L.
Signaling through the Toll receptor is required for dorsal/ventral polarity in Drosophila embryos, and also plays an evolutionarily conserved role in the immune response. Upon ligand binding, Toll...
Prasad, Jayendra, Colwill, Karen, Pawson, Tony, Manley, James L.
The splicing of mammalian mRNA precursors requires both protein phosphorylation and dephosphorylation, likely involving modification of members of the SR protein family of splicing factors. Several...
Complex Protein Interactions within the Human Polyadenylation Machinery Identify a Novel Component
Takagaki, Yoshio, Manley, James L.
Polyadenylation of mRNA precursors is a two-step reaction requiring multiple protein factors. Cleavage stimulation factor (CstF) is a heterotrimer necessary for the first step, endonucleolytic...
We have employed gene targeting coupled with conditional expression to construct a chicken DT40 cell line in which a tetracycline (Tet)-repressible promoter is exclusively responsible for expression...
Poly(A) Polymerase Phosphorylation Is Dependent on Novel Interactions with Cyclins
Bond, Gareth L., Prives, Carol, Manley, James L.
We have previously shown that poly(A) polymerase (PAP) is negatively regulated by cyclin B-cdc2 kinase hyperphosphorylation in the M phase of the cell cycle. Here we show that cyclin B1 binds PAP...
Um, Moonkyoung, Yamauchi, Jun, Kato, Shigeaki, Manley, James L.
TATA-binding protein (TBP) is a key general transcription factor required for transcription by all three nuclear RNA polymerases. Although it has been intensively analyzed in vitro and in...
Topalian, Suzanne L., Kaneko, Syuzo, Gonzales, Monica I., Bond, Gareth L., Ward, Yvona, Manley, James L.
Poly(A) polymerase (PAP) plays an essential role in polyadenylation of mRNA precursors, and it has long been thought that mammalian cells contain only a single PAP gene. We describe here the...
Hatton, Lee S., Eloranta, Jyrki J., Figueiredo, Luisa M., Takagaki, Yoshio, Manley, James L., O’Hare, Kevin
During mRNA 3′ end formation, cleavage stimulation factor (CstF) binds to a GU-rich sequence downstream from the polyadenylation site and helps to stabilise the binding of cleavage-polyadenylation...
The Drosophila homeodomain protein Even-skipped (Eve) is a transcriptional repressor, and previous studies have suggested that it functions by interfering with the basal transcription machinery. Here...
Deregulation of Poly(A) Polymerase Interferes with Cell Growth
Zhao, Wenqing, Manley, James L.
Vertebrate poly(A) polymerase (PAP) contains a catalytic domain and a C-terminal Ser-Thr-rich regulatory region. Consensus and nonconsensus cyclin-dependent kinase (cdk) sites are conserved in the...
Requirements of the RNA Polymerase II C-Terminal Domain for Reconstituting Pre-mRNA 3′ Cleavage
Ryan, Kevin, Murthy, Kanneganti G. K., Kaneko, Syuzo, Manley, James L.
RNA polymerase II (RNAP II) has previously been shown to be required for the pre-mRNA polyadenylation cleavage reaction in vitro. This activity was found to reside solely in the C-terminal domain...
ASAP, a Novel Protein Complex Involved in RNA Processing and Apoptosis
Schwerk, Christian, Prasad, Jayendra, Degenhardt, Kurt, Erdjument-Bromage, Hediye, White, Eileen, Tempst, Paul, ...
Different isoforms of a protein complex termed the apoptosis- and splicing-associated protein (ASAP) were isolated from HeLa cell extract. ASAP complexes are composed of the polypeptides SAP18 and...
Lemaire, Raphael, Prasad, Jayendra, Kashima, Tsuyoshi, Gustafson, Jennifer, Manley, James L., Lafyatis, Robert
Pre-mRNA splicing is a widely used regulatory mechanism for controlling gene expression, and a family of conserved proteins, SR proteins, participate in both constitutive and alternative splicing....
Regulation and Substrate Specificity of the SR Protein Kinase Clk/Sty
Prasad, Jayendra, Manley, James L.
SR proteins constitute a family of splicing factors that play key roles in both constitutive and regulated splicing in metazoan organisms. The proteins are extensively phosphorylated, and kinases...
Gene-specific transcriptional activation is a multistep process that requires numerous protein factors and DNA elements, including enhancers and the core promoter. To investigate the roles of core...
Pin1 modulates the structure and function of human RNA polymerase II
Xu, Yu-Xin, Hirose, Yutaka, Zhou, Xiao Zhen, Lu, Kun Ping, Manley, James L.
The C-terminal domain of the RNA polymerase (RNAP) II largest subunit (CTD) plays critical roles both in transcription of mRNA precursors and in the processing reactions needed to form mature mRNAs....
High mobility group protein-1 (HMG-1) is a unique activator of p53
Jayaraman, Lata, Moorthy, Narayani Chandra, Murthy, Kanneganti G.K., Manley, James L., Bustin, Michael, Prives, Carol
The binding of p53 protein to DNA is stimulated by its interaction with covalent as well as noncovalent modifiers. We report the identification of a factor from HeLa nuclear extracts that activates...
Phosphorylated RNA polymerase II stimulates pre-mRNA splicing
Hirose, Yutaka, Tacke, Roland, Manley, James L.
RNA polymerase II (RNAP II) is responsible for transcription of mRNA precursors in eukaryotic cells. Recent studies, however, have suggested that RNAP II also participates in subsequent RNA...
Wang, Jin, Xiao, Shou-Hua, Manley, James L.
The SR proteins constitute a family of splicing factors, highly conserved in metazoans, that contain one or two amino-terminal RNA-binding domains (RBDs) and a region enriched in arginine/serine...
Moreira, Alexandra, Takagaki, Yoshio, Brackenridge, Simon, Wollerton, Matthew, Manley, James L., Proudfoot, Nicholas J.
The poly(A) signal of the C2 complement gene is unusual in that it possesses an upstream sequence element (USE) required for full activity in vivo. We describe here in vitro experiments demonstrating...
Millhouse, Scott, Manley, James L.
RNA polymerase II, and specifically the C-terminal domain (CTD) of its largest subunit, has been demonstrated to play important roles in capping, splicing, and 3′ processing of mRNA precursors. But...
The transcriptional coactivator PC4/Sub1 has multiple functions in RNA polymerase II transcription
Transcription and processing of mRNA precursors are coordinated events that require numerous complex interactions to ensure that they are successfully executed. We described previously an unexpected...
BRCA1/BARD1 inhibition of mRNA 3′ processing involves targeted degradation of RNA polymerase II
Kleiman, Frida E., Wu-Baer, Foon, Fonseca, Danae, Kaneko, Syuzo, Baer, Richard, Manley, James L.
Mammalian cells exhibit a complex response to DNA damage. The tumor suppressor BRCA1 and associated protein BARD1 are thought to play an important role in this response, and our previous work...
Multiple Properties of the Splicing Repressor SRp38 Distinguish It from Typical SR Proteins
Shin, Chanseok, Kleiman, Frida E., Manley, James L.
The SR protein SRp38 is a general splicing repressor that is activated by dephosphorylation during mitosis and in response to heat shock. Here we describe experiments that provide insights into the...
Li, Xialu, Wang, Jin, Manley, James L.
ASF/SF2 is an SR protein splicing factor that participates in constitutive and alternative pre-mRNA splicing and is essential for cell viability. Using a genetically modified chicken B-cell line,...
Hsp27 Enhances Recovery of Splicing as well as Rephosphorylation of SRp38 after Heat Shock
Marin-Vinader, Laura, Shin, Chanseok, Onnekink, Carla, Manley, James L., Lubsen, Nicolette H.
A heat stress causes a rapid inhibition of splicing. Exogenous expression of Hsp27 did not prevent that inhibition but enhanced the recovery of splicing afterward. Another small heat shock protein,...
Characterization of the catalytic activity of U2 and U6 snRNAs
VALADKHAN, SABA, MANLEY, JAMES L.
Removal of introns from pre-messenger RNAs in eukaryotes is carried out by the spliceosome, an assembly of a large number of proteins and five small nuclear RNAs (snRNAs). We showed previously that...
Evidence that polyadenylation factor CPSF-73 is the mRNA 3′ processing endonuclease
RYAN, KEVIN, CALVO, OLGA, MANLEY, JAMES L.
Generation of the polyadenylated 3′ end of an mRNA requires an endonucleolytic cleavage followed by synthesis of the poly(A) tail. Despite the seeming simplicity of the reaction, more than a dozen...
Regulation of Plant Developmental Processes by a Novel Splicing Factor
Ali, Gul Shad, Palusa, Saiprasad G., Golovkin, Maxim, Prasad, Jayendra, Manley, James L., Reddy, Anireddy S.N.
Serine/arginine-rich (SR) proteins play important roles in constitutive and alternative splicing and other aspects of mRNA metabolism. We have previously isolated a unique plant SR protein (SR45)...
An intronic element contributes to splicing repression in spinal muscular atrophy
Kashima, Tsuyoshi, Rao, Nishta, Manley, James L.
The neurodegenerative disease spinal muscular atrophy is caused by mutation of the survival motor neuron 1 (SMN1) gene. SMN2 is a nearly identical copy of SMN1 that is unable to prevent disease,...
Kaneko, Syuzo, Rozenblatt-Rosen, Orit, Meyerson, Matthew, Manley, James L.
Termination of RNA polymerase II transcription frequently requires a poly(A) signal and cleavage/polyadenylation factors. Recent work has shown that degradation of the downstream cleaved RNA by the...
Concurrent splicing and transcription are not sufficient to enhance splicing efficiency
Lazarev, Denis, Manley, James L.
The concept of a tight integration of transcription and splicing of mRNA precursors has been supported with increasing evidence in recent years. However, the mechanism and functional consequences of...
Sumoylation Modulates the Assembly and Activity of the Pre-mRNA 3′ Processing Complex▿ †
Vethantham, Vasupradha, Rao, Nishta, Manley, James L.
Eukaryotic pre-mRNA 3′-end formation is catalyzed by a complex set of factors that must be intricately regulated. In this study, we have discovered a novel role for the small ubiquitin-like...
Bush, Stephanie D., Richard, Patricia, Manley, James L.
We previously showed that reduced intracellular levels of the TATA binding protein (TBP), brought about by tbp heterozygosity in DT40 cells, resulted in a mitotic delay reflecting reduced expression...
Human capping enzyme promotes formation of transcriptional R loops in vitro
Kaneko, Syuzo, Chu, Chun, Shatkin, Aaron J., Manley, James L.
Cap formation is the first step of pre-mRNA processing in eukaryotic cells. Immediately after transcription initiation, capping enzyme (CE) is recruited to RNA polymerase II (Pol II) by the...
The 3′ processing factor CstF functions in the DNA repair response
Mirkin, Nurit, Fonseca, Danae, Mohammed, Samia, Cevher, Murat A., Manley, James L., Kleiman, Frida E.
Following DNA damage, mRNA levels decrease, reflecting a coordinated interaction of the DNA repair, transcription and RNA processing machineries. In this study, we provide evidence that transcription...
Pin1 modulates RNA polymerase II activity during the transcription cycle
The C-terminal domain of the RNA polymerase (RNAP) II largest subunit (CTD) plays a critical role in coordinating multiple events in pre-mRNA transcription and processing. Previously we reported that...
Protein-free spliceosomal snRNAs catalyze a reaction that resembles the first step of splicing
Valadkhan, Saba, Mohammadi, Afshin, Wachtel, Chaim, Manley, James L.
Splicing of introns from mRNA precursors is a two-step reaction performed by the spliceosome, an immense cellular machine consisting of over 200 different proteins and five small RNAs (snRNAs). We...
The RNA binding protein RNPS1 alleviates ASF/SF2 depletion-induced genomic instability
Li, Xialu, Niu, Tianhui, Manley, James L.
Formation of transcription-induced R-loops poses a critical threat to genomic integrity throughout evolution. We have recently shown that the SR protein ASF/SF2 prevents R-loop formation in...
Sumoylation regulates multiple aspects of mammalian poly(A) polymerase function
Vethantham, Vasupradha, Rao, Nishta, Manley, James L.
The addition of the poly(A) tail to the ends of eukaryotic mRNAs is catalyzed by poly(A) polymerase (PAP). PAP activity is known to be highly regulated, for example, by alternative splicing and...
The tumor suppressor Cdc73 functionally associates with CPSF and CstF 3′ mRNA processing factors
Rozenblatt-Rosen, Orit, Nagaike, Takashi, Francis, Joshua M., Kaneko, Syuzo, Glatt, Karen A., Hughes, Christina M., ...
The CDC73 tumor suppressor gene is mutationally inactivated in hereditary and sporadic parathyroid tumors. Its product, the Cdc73 protein, is a component of the RNA polymerase II and...
The use of simple model systems to study spliceosomal catalysis
Valadkhan, Saba, Manley, James L.
Since direct analysis of many aspects of spliceosomal function is greatly hindered by the daunting complexity of the spliceosome, the development of functionally validated simple model systems can be...
Sub1 Functions in Osmoregulation and in Transcription by both RNA Polymerases II and III▿ †
Rosonina, Emanuel, Willis, Ian M., Manley, James L.
Sub1 is implicated in transcriptional activation, elongation, and mRNA 3′-end formation in budding yeast. To gain more insight into its function, we performed a synthetic genetic array screen with...