U. L. RajBhandary

Publication List Details

Period

1974 - 2004

Number

135

Co-Authors

Mycobacterium tuberculosis Rv2118c codes for a single-component homotetrameric m1A58 tRNA methyltransferase (2004)

Varshney, U., Ramesh, V., Madabushi, A., Gaur, R., Subramanya, H. S., RajBhandary, U. L.

Modified nucleosides in tRNAs play important roles in tRNA structure, biosynthesis and function, and serve as crucial determinants of bacterial growth and virulence. In the yeast Saccharomyces...

Chemical reactivity of E. coli 5S RNA in situ in the 50S ribosomal subunit (1983)

Silberklang, M., RajBhandary, U. L., Luck, A., Erdmann, V. A.

E.coli 50S ribosomal subunits were reacted with monoper-phthalic acid under conditions in which non-base paired adenines are modified to their 1-N-oxides. 5S RNA was isolated from such chemically...

The nucleotide sequence of blue-green algae phenylalanine-tRNA and the evolutionary origin of chloroplasts (1982)

Hecker, L.J., Bamett, W.E., Lin, F.K., Furr, T.D., Heckman, J.E., RajBhandary, U.L., ...

Phenylalanine tRNA from the blue-green alga, Agmenellum quadruplicatum, has been purified to homogeneity. The nucleotide sequence of this tRNA was determined to be:...

The nucleotide sequence of the initiator tRNA from Drosophila melanogaster (1979)

Silverman, S., Heckman, J., Cowling, G.J., Delaney, A.D., Dunn, R.J., Gillani, I.C., ...

The nucleotide sequence of Drosqphila melanogaster methionine tRNAi was determined to be:...

The nucleotide sequence of formylmethionine tRNA from Mycoplasma mycoides sp. capri (1978)

Walker, R.T., RajBhandary, U.L.

The nucleotide sequence of Mycoplasma mycoides sp. capri PG3 formylmethionine tRNA has been determined, using in vitro labeling techniques, to be...

The use of nuclease P1 in sequence analysis of end group labeled RNA (1977)

Silberklang, M., Gillum, A.M., Rajbhandary, U.L.

A method is described for the direct sequence analysis 20–25 nucleotides from the terinini of 5′- or 3′-end-group [32P] labeled RNA. The method involves partial endonucleolytic digestion of the...

Nucleotide sequence of Neurospora crassa cytoplasmic initiator tRNA (1977)

Gillum, A.M., Hecker, L.I., Silberklang, M., Schwartzbach, S.D., RajBhandary, U.L., Barnett, W.Edgar

Initiator methionine tRNA from the cytoplasm of Neurospora crassa has been purified and sequenced. The sequence is: pAGCUGCAUm1$${\hbox{ GGCGCAGCGGAAGCGCm }}_{\hbox{ 2 }}^{\hbox{ 2 }}$$2GCGY*CUCAUt6...

Formylatable methionine transfer RNA from Mycoplasma: purification and comparison of partial nucleotide sequences with those of other prokaryotic initiator tRNAs (1975)

Walker, R.T., RajBhandary, U.L.

The major species of the formylatable methionine tRNA from Mycoiplasma mycoides var capri has been purified. The 5′- and 3′- terminal sequences of the purified tRNA are pC-G-and C-A-A-C-C-AOH,...

2'-O-Methyl ribothymidine: a component of rabbit liver lysine transfer RNA (1974)

Gross, H.J., Simsek, M., Raba, M., Limburg, K., Heckman, J., RajBhandary, U.L.

One of the lysine transfer RNAs of rabbit liver is shown to contain 2′-O-methyl ribothymidine in place of ribothymidine. This represents the first demonstration of the presence of 2′-O- methyl...

NMR analysis of tRNA acceptor stem microhelices: discriminator base change affects tRNA conformation at the 3' end.

Puglisi, E V, Puglisi, J D, Williamson, J R, RajBhandary, U L

An important step in initiation of protein synthesis in Escherichia coli is the specific formylation of the initiator methionyl-tRNA (Met-tRNA) by Met-tRNA transformylase. The determinants for...

From elongator tRNA to initiator tRNA.

Varshney, U, Lee, C P, RajBhandary, U L

We show that the two most important properties needed for a tRNA to function in initiation in Escherichia coli are its ability to be formylated and its ability to bind to the ribosomal P site. This...

The discriminator base influences tRNA structure at the end of the acceptor stem and possibly its interaction with proteins.

Lee, C P, Mandal, N, Dyson, M R, RajBhandary, U L

For many tRNAs, the discriminator base preceding the CCA sequence at the 3' end is important for aminoacylation. We show that the discriminator base influences the stability of the 1.72 base pair...

Striking effects of coupling mutations in the acceptor stem on recognition of tRNAs by Escherichia coli Met-tRNA synthetase and Met-tRNA transformylase.

Lee, C P, Dyson, M R, Mandal, N, Varshney, U, Bahramian, B, RajBhandary, U L

We measured kinetic parameters in vitro and directly analyzed aminoacylation and formylation levels in vivo to study recognition of Escherichia coli initiator tRNA mutants by E. coli Met-tRNA...

Expression of the bacterioopsin gene in Halobacterium halobium using a multicopy plasmid.

Krebs, M P, Hauss, T, Heyn, M P, RajBhandary, U L, Khorana, H G

Bacteriorhodopsin (bR) was expressed in Halobacterium halobium by using a multicopy plasmid containing the bop gene. The plasmid contains pGRB1, a 1.8-kilobase-pair plasmid; a 70-base-pair fragment...

Mutants of Escherichia coli initiator tRNA that suppress amber codons in Saccharomyces cerevisiae and are aminoacylated with tyrosine by yeast extracts.

Lee, C P, RajBhandary, U L

We recently described mutants of Escherichia coli initiator tRNA that suppress amber termination codons (UAG) in E. coli. These mutants have changes in the anticodon sequence (CAU----CUA) that allow...

Initiation of protein synthesis from a termination codon.

Varshney, U, RajBhandary, U L

We show that the amber termination codon UAG can initiate protein synthesis in Escherichia coli. We mutated the initiation codon AUG of the chloramphenicol acetyltransferase (CAT) gene to UAG...

Structural studies on tRNA acceptor stem microhelices: exchange of the discriminator base A73 for G in human tRNALeu switches the acceptor specificity from leucine to serine possibly by decreasing the stability of the terminal G1-C72 base pair.

Metzger, A U, Heckl, M, Willbold, D, Breitschopf, K, RajBhandary, U L, Rösch, P, ...

Correct recognition of transfer RNAs (tRNAs) by aminoacyl-tRNA synthetases (aaRS) is crucial to the maintenance of translational fidelity. The discriminator base A73 in human tRNALeuis critical for...

An anticodon sequence mutant of Escherichia coli initiator tRNA: possible importance of a newly acquired base modification next to the anticodon on its activity in initiation.

Mangroo, D, Limbach, P A, McCloskey, J A, RajBhandary, U L

Initiator tRNAs from eubacteria and chloroplasts lack a base modification next to the anticodon. This is in contrast to virtually all other tRNAs from these sources. We show that a mutant Escherichia...

Developmental regulation of the gene for formate dehydrogenase in Neurospora crassa.

Chow, C M, RajBhandary, U L

We have isolated and characterized a gene, fdh, from Neurospora crassa which is developmentally regulated and which produces formate dehydrogenase activity when expressed in Escherichia coli. The...

Role of methionine and formylation of initiator tRNA in initiation of protein synthesis in Escherichia coli.

Varshney, U, RajBhandary, U L

We showed recently that a mutant of Escherichia coli initiator tRNA with a CAU-->CUA anticodon sequence change can initiate protein synthesis from UAG by using formylglutamine instead of...

Escherichia coli B lacks one of the two initiator tRNA species present in E. coli K-12.

Mandal, N, RajBhandary, U L

We show that the metY locus which specifies tRNA(2fMet) in Escherichia coli K-12 specifies tRNA(1fMet) in E. coli B. This conclusion is based on results of Southern blot analysis of E. coli B and...

Amber suppression in mammalian cells dependent upon expression of an Escherichia coli aminoacyl-tRNA synthetase gene.

Drabkin, H J, Park, H J, RajBhandary, U L

As an approach to inducible suppression of nonsense mutations in mammalian and in higher eukaryotic cells, we have analyzed the expression of an Escherichia coli glutamine-inserting amber suppressor...

The A1 x U72 base pair conserved in eukaryotic initiator tRNAs is important specifically for binding to the eukaryotic translation initiation factor eIF2.

Farruggio, D, Chaudhuri, J, Maitra, U, RajBhandary, U L

The formation of a specific ternary complex between eukaryotic initiation factor 2 (eIF2), the initiator methionyl-tRNA (Met-tRNA), and GTP is a critical step in translation initiation in the...

Functional suppression in mammalian cells of nonsense mutations in the herpes simplex virus thymidine kinase gene by suppressor tRNA genes.

Summers, W P, Summers, W C, Laski, F A, RajBhandary, U L, Sharp, P A

A nonsense mutation (UAG) in the thymidine kinase gene of herpes simplex virus type 1 can be suppressed in vivo to produce active thymidine kinase by prior infection with a defective simian virus 40...

Construction, stable transformation, and function of an amber suppressor tRNA gene in Drosophila melanogaster.

Laski, F A, Ganguly, S, Sharp, P A, RajBhandary, U L, Rubin, G M

Drosophila melanogaster strains with a stably incorporated amber suppressor tRNA gene have been generated. A tRNATyr gene was site specifically mutated to produce an anticodon sequence that...

Mutants of Escherichia coli formylmethionine tRNA: a single base change enables initiator tRNA to act as an elongator in vitro.

Seong, B L, RajBhandary, U L

We show that the absence of a Watson-Crick base pair at the end of the amino acid acceptor stem, which is a hallmark of all prokaryotic initiator tRNAs, is one of the key features that prevents them...

Escherichia coli formylmethionine tRNA: mutations in GGGCCC sequence conserved in anticodon stem of initiator tRNAs affect initiation of protein synthesis and conformation of anticodon loop.

Seong, B L, RajBhandary, U L

We have generated mutants of Escherichia coli formylmethionine initiator tRNA in which one, two, and all three G X C base pairs in the GGGCCC sequence in the anticodon stem are changed to those found...

Bacteriorhodopsin: partial sequence of mRNA provides amino acid sequence in the precursor region.

Chang, S H, Majumdar, A, Dunn, R, Makabe, O, RajBhandary, U L, Khorana, H G, ...

mRNA for bacteriorhodopsin from Halobacterium halobium has been partially purified. By using this mRNA as template in the presence of reverse transcriptase RNA-dependent DNA nucleotidyltransferase...

Site-specific mutagenesis on cloned DNAs: generation of a mutant of Escherichia coli tyrosine suppressor tRNA in which the sequence G-T-T-C corresponding to the universal G-T-pseudouracil-C sequence of tRNAs is changed to G-A-T-C.

Kudo, I, Leineweber, M, RajBhandary, U L

We have cloned the Escherichia coli tyrosine-inserting amber suppressor tRNA gene into the recombinant single-strand phage M12mp3. By using the M13mp3SuIII+ recombinant phage DNA as template and an...

Nucleotide sequence of wheat germ cytoplasmic initiator methionine transfer ribonucleic acid.

Ghosh, H P, Ghosh, K, Simsek, M, RajBhandary, U L

The primary sequence of wheat germ initiator tRNA has been determined using in vitro labelling techniques. The sequence is: pAUCAGAGUm1Gm2GCGCAG CGGAAGCGUm2GG psi GGGCCCAUt6AACCCACAGm7GDm5Cm5CCAGGA...

Expression of a X. laevis tRNATyr gene in mammalian cells.

Laski, F A, Alzner-DeWeerd, B, RajBhandary, U L, Sharp, P A

Expression of a X. laevis tRNATyr gene has been studied in mammalian cells. This tRNATyr gene has a 13 base intervening sequence adjacent to its anticodon. A fragment containing the tRNATyr gene was...

The nucleotide sequence of phenylalanine tRNA from the cytoplasm of Neurospora crassa.

Alzner-DeWeerd, B, Hecker, L I, Barnett, W E, RajBhandary, U L

The phenylalanine tRNA from the cytoplasm of Neurospora crassa has been purified and sequenced. The sequence is: pGCGGGUUUAm2GCUCA (N)...

Chemical reactivity of E. coli 5S RNA in situ in the 50S ribosomal subunit.

Silberklang, M, RajBhandary, U L, Lück, A, Erdmann, V A

E. coli 50S ribosomal subunits were reacted with monoperphthalic acid under conditions in which non-base paired adenines are modified to their 1-N-oxides. 5S RNA was isolated from such chemically...

The nucleotide sequence of blue-green algae phenylalanine-tRNA and the evolutionary origin of chloroplasts.

Hecker, L I, Barnett, W E, Lin, F K, Furr, T D, Heckman, J E, RajBhandary, U L, ...

Phenylalanine tRNA from the blue-green alga, Agmenellum quadruplicatum, has been purified to homogeneity. The nucleotide sequence of this tRNA was determined to be: (see tests) Comparisons of the...

The nucleotide sequence of Euglena cytoplasmic phenylalanine transfer RNA. Evidence for possible classifications of Euglena among the animal rather than the plant kingdom.

Chang, S H, Hecker, L I, Brum, C K, Schnabel, J J, Heckman, J E, Silberklang, M, ...

The nucleotide sequence of cytoplasmic phenylalanine tRNA from Euglena gracilis has been elucidated using procedures described previously for the corresponding chloroplastic tRNA [Cell, 9, 717...

The nucleotide sequence of the initiator tRNA from Drosophila melanogaster.

Silverman, S, Heckman, J, Cowling, G J, Delaney, A D, Dunn, R J, Gillam, I C, ...

The nucleotide sequence of Drosophila melanogaster methionine tRNAi was determined to be:...

Sequence analysis of 5'[32P] labeled mRNA and tRNA using polyacrylamide gel electrophoresis.

Lockard, R E, Alzner-Deweerd, B, Heckman, J E, MacGee, J, Tabor, M W, RajBhandary, U L

Sequence analysis of 5'-[32P] labeled tRNA and eukaryotic mRNA using an adaptation of a method recently described by Donis-Keller, Maxam and Gilbert for mapping guanines, adenines and pyrimidines...

The nucleotide sequence of formylmethionine tRNA from Mycoplasma mycoides sp. capri.

Walker, R T, RajBhandary, U L

The nucleotide sequence of Mycoplasma mycoides sp. capri PG3 formylmethionine tRNA has been determined, using in vitro labeling techniques, to be pC-G-C-G-G-G-G-s4U-A-G-A-G-C-A-G-U-D...

Formylatable methionine transfer RNA from Mycoplasma: purification and comparison of partial nucleotide sequences with those of other prokaryotic initiator tRNAs.

Walker, R T, RajBhandary, U L

The major species of the formylatable methionine tRNA from Mycoplasma mycoides var capri has been purified. The 5'- and 3'-terminal sequences of the purified tRNA are pC-G- and C-A-A-C-C-AOH,...

The use of nuclease P1 in sequence analysis of end group labeled RNA.

Silberklang, M, Gillum, A M, RajBhandary, U L

A method is described for the direct sequence analysis of 20-25 nucleotides from the termini of 5'- or 3'-end-group [32P] labeled RNA. The method involves partial endonucleolytic digestion of the...

Nucleotide sequence of Neurospora crassa cytoplasmic initiator tRNA.

Gillum, A M, Hecker, L I, Silberklang, M, Schwartzbach, S D, RajBhandary, U L, Barnett, W E

Initiator methionine tRNA from the cytoplasm of Neurospora crassa has been purified and sequenced. The sequence is: pAGCUGCAUm1GGCGCAGCGGAAGCGCM22GCY*GGGCUCAUt6AACCCGGAGm7GU (or D) -...

2′-O-Methyl ribothymidine: a component of rabbit liver lysine transfer RNA

Gross, H.J., Simsek, M., Raba, M., Limburg, K., Heckman, J., RajBhandary, U.L.

One of the lysine transfer RNAs of rabbit liver is shown to contain 2′-O-methyl ribothymidine in place of ribothymidine. This represents the first demonstration of the presence of 2′-O-methyl...

An amber suppressor tRNA gene derived by site-specific mutagenesis: cloning and function in mammalian cells.

Laski, F A, Belagaje, R, RajBhandary, U L, Sharp, P A

We describe the synthesis, cloning, expression, and in vivo function of a suppressor tRNA gene in mammalian cells. By using "primer-directed mutagenesis" on a Xenopus laevis tyrosine tRNA gene cloned...

A transposable element from Halobacterium halobium which inactivates the bacteriorhodopsin gene.

Simsek, M, DasSarma, S, RajBhandary, U L, Khorana, H G

We describe the characterization of a transposable element from an archaebacterium. The bacteriorhodopsin genes from the wild-type and two mutant Halobacterium halobium strains have been cloned as...

The bacteriorhodopsin gene.

Dunn, R, McCoy, J, Simsek, M, Majumdar, A, Chang, S H, Rajbhandary, U L, ...

The bacteriorhodopsin gene has been identified in a 5.3-kilobase restriction endonuclease fragment isolated from Halobacterium halobium DNA, using a cloned cDNA fragment as the probe. Of the 1229...

Expression and function of a human initiator tRNA gene in the yeast Saccharomyces cerevisiae.

Francis, M A, Rajbhandary, U L

We showed previously that the human initiator tRNA gene, in the context of its own 5'- and 3'-flanking sequences, was not expressed in Saccharomyces cerevisiae. Here we show that switching its...

Cytochrome oxidase subunit V gene of Neurospora crassa: DNA sequences, chromosomal mapping, and evidence that the cya-4 locus specifies the structural gene for subunit V.

Sachs, M S, Bertrand, H, Metzenberg, R L, RajBhandary, U L

The sequences of cDNA and genomic DNA clones for Neurospora cytochrome oxidase subunit V show that the protein is synthesized as a 171-amino-acid precursor containing a 27-amino-acid N-terminal...

Saccharomyces cerevisiae SUP53 tRNA gene transcripts are processed by mammalian cell extracts in vitro but are not processed in vivo.

Ganguly, S, Sharp, P A, RajBhandary, U L

We describe the results of our studies of expression of a Saccharomyces cerevisiae amber suppressor tRNA(Leu) gene (SUP53) in mammalian cells in vivo and in cell extracts in vitro. Parallel studies...

Nuclear gene for mitochondrial leucyl-tRNA synthetase of Neurospora crassa: isolation, sequence, chromosomal mapping, and evidence that the leu-5 locus specifies structural information.

Chow, C M, Metzenberg, R L, Rajbhandary, U L

We have isolated and characterized the nuclear gene for the mitochondrial leucyl-tRNA synthetase (LeuRS) of Neurospora crassa and have established that a defect in this structural gene is responsible...

Regulation of the nuclear genes encoding the cytoplasmic and mitochondrial leucyl-tRNA synthetases of Neurospora crassa.

Chow, C M, Rajbhandary, U L

We show that the nuclear genes for the cytoplasmic and mitochondrial leucyl-tRNA synthetase (LeuRS) of Neurospora crassa are distinct in their encoded proteins, codon usage, mRNA levels, and...

Introduction of UAG, UAA, and UGA nonsense mutations at a specific site in the Escherichia coli chloramphenicol acetyltransferase gene: use in measurement of amber, ochre, and opal suppression in mammalian cells.

Capone, J P, Sedivy, J M, Sharp, P A, RajBhandary, U L

We have used oligonucleotide-directed site-specific mutagenesis to convert serine codon 27 of the Escherichia coli chloramphenicol acetyltransferase (cat) gene to UAG, UAA, and UGA nonsense codons....

Mycobacterium tuberculosis Rv2118c codes for a single-component homotetrameric m1A58 tRNA methyltransferase

Varshney, U., Ramesh, V., Madabushi, A., Gaur, R., Subramanya, H. S., RajBhandary, U. L.

Modified nucleosides in tRNAs play important roles in tRNA structure, biosynthesis and function, and serve as crucial determinants of bacterial growth and virulence. In the yeast Saccharomyces...

Interesting and unusual features in the sequence of Neurospora crassa mitochondrial tyrosine transfer RNA.

Heckman, J E, Alzner-Deweerd, B, RajBhandary, U L

The mitochondrial tyrosine tRNA from Neurospora crassa has been sequenced and found to have several interesting features: (i) It resembles prokaryotic rather than eukaryotic tyrosine tRNAs in that it...

High-frequency spontaneous mutation in the bacterio-opsin gene in Halobacterium halobium is mediated by transposable elements.

DasSarma, S, RajBhandary, U L, Khorana, H G

We have recently characterized a transposable element, ISH1, which inactivates the bacterio-opsin (BO) gene in two purple membrane-deficient (Pum-) mutants of Halobacterium halobium. Examination of...

Nucleotide sequence of 5' terminus of alfalfa mosaic virus RNA 4 leading into coat protein cistron.

Koper-Zwarthoff, E C, Lockard, R E, Alzner-deWeerd, B, RajBhandary, U L, Bol, J F

The sequence of the 5'-terminal 74 nucleotides of alfalfa mosaic virus RNA 4, the mRNA for the viral coat protein, has been deduced by using various new techniques for labeling the RNA at the 5' end...

Ribosome-initiator tRNA complex as an intermediate in translation initiation in Escherichia coli revealed by use of mutant initiator tRNAs and specialized ribosomes.

Wu, X Q, Iyengar, P, RajBhandary, U L

For functional studies of mutant Escherichia coli initiator tRNAs in vivo, we previously described a strategy based on the use of tRNA genes carrying an anticodon sequence change from CAU to CUA...

RNA processing in Neurospora crassa mitochondria: use of transfer RNA sequences as signals.

Breitenberger, C A, Browning, K S, Alzner-DeWeerd, B, RajBhandary, U L

We have used RNA gel transfer hybridization, S1 nuclease mapping and primer extension to analyze transcripts derived from several genes in Neurospora crassa mitochondria. The transcripts studied...

Amber, ochre and opal suppressor tRNA genes derived from a human serine tRNA gene.

Capone, J P, Sharp, P A, RajBhandary, U L

Amber, ochre and opal suppressor tRNA genes have been generated by using oligonucleotide directed site-specific mutagenesis to change one or two nucleotides in a human serine tRNA gene. The amber and...

Synthesis of an ochre suppressor tRNA gene and expression in mammalian cells.

Laski, F A, Belagaje, R, Hudziak, R M, Capecchi, M R, Norton, G P, Palese, P, ...

We have used site-specific mutagenesis to change the anticodon of a Xenopus laevis tyrosine tRNA gene so that it would recognize ochre codons. This tRNA gene is expressed when amplified in monkey...

The E35 stopper mutant of Neurospora crassa: precise localization of deletion endpoints in mitochondrial DNA and evidence that the deleted DNA codes for a subunit of NADH dehydrogenase.

De Vries, H, Alzner-DeWeerd, B, Breitenberger, C A, Chang, D D, De Jonge, J C, RajBhandary, U L

Two types of defective mitochondrial DNA molecules with large deletions (5 kbp and 40 kbp) have previously been identified in the stopper mutant, E35, of Neurospora crassa. The junction fragments...

Role of the three consecutive G:C base pairs conserved in the anticodon stem of initiator tRNAs in initiation of protein synthesis in Escherichia coli.

Mandal, N, Mangroo, D, Dalluge, J J, McCloskey, J A, Rajbhandary, U L

The three consecutive G:C base pairs, G29:C41, G30:C40, and G31:C39, are conserved in the anticodon stem of virtually all initiator tRNAs from eubacteria, eukaryotes, and archaebacteria. We show that...

Intragenic suppression in tRNA: evidence for crosstalk between the D and the T stems.

Ramesh, V, Varshney, U, Rajbhandary, U L

We showed previously that introduction of two of the three unique features of Escherichia coli initiator tRNA onto an elongator methionine tRNA conferred significant activity in initiation....

Altered discrimination of start codons and initiator tRNAs by mutant initiation factor 3.

O'Connor, M, Gregory, S T, Rajbhandary, U L, Dahlberg, A E

IF3 is essential for ensuring the fidelity of the initiation step of translation in bacterial cells. Mutations at residues R99 and R131 in the C-terminal domain of the factor have previously been...

Cytoplasmic Leucyl-Trna Synthetase of Neurospora Crassa Is Not Specified by the Leu-5 Locus

Benarous, R., Chow, C. M., RajBhandary, U. L.

We generated a λgt11 Neurospora crassa cDNA library and screened the library for the cytoplasmic leucyl-tRNA synthetase (cyto LeuRS) clones using cyto LeuRS specific antibody. Two clones, λNCLRSC1...

NMR analysis of tRNA acceptor stem microhelices: discriminator base change affects tRNA conformation at the 3' end.

Puglisi, E V, Puglisi, J D, Williamson, J R, RajBhandary, U L

An important step in initiation of protein synthesis in Escherichia coli is the specific formylation of the initiator methionyl-tRNA (Met-tRNA) by Met-tRNA transformylase. The determinants for...

From elongator tRNA to initiator tRNA.

Varshney, U, Lee, C P, RajBhandary, U L

We show that the two most important properties needed for a tRNA to function in initiation in Escherichia coli are its ability to be formylated and its ability to bind to the ribosomal P site. This...

The discriminator base influences tRNA structure at the end of the acceptor stem and possibly its interaction with proteins.

Lee, C P, Mandal, N, Dyson, M R, RajBhandary, U L

For many tRNAs, the discriminator base preceding the CCA sequence at the 3' end is important for aminoacylation. We show that the discriminator base influences the stability of the 1.72 base pair...

Striking effects of coupling mutations in the acceptor stem on recognition of tRNAs by Escherichia coli Met-tRNA synthetase and Met-tRNA transformylase.

Lee, C P, Dyson, M R, Mandal, N, Varshney, U, Bahramian, B, RajBhandary, U L

We measured kinetic parameters in vitro and directly analyzed aminoacylation and formylation levels in vivo to study recognition of Escherichia coli initiator tRNA mutants by E. coli Met-tRNA...

Expression of the bacterioopsin gene in Halobacterium halobium using a multicopy plasmid.

Krebs, M P, Hauss, T, Heyn, M P, RajBhandary, U L, Khorana, H G

Bacteriorhodopsin (bR) was expressed in Halobacterium halobium by using a multicopy plasmid containing the bop gene. The plasmid contains pGRB1, a 1.8-kilobase-pair plasmid; a 70-base-pair fragment...

Mutants of Escherichia coli initiator tRNA that suppress amber codons in Saccharomyces cerevisiae and are aminoacylated with tyrosine by yeast extracts.

Lee, C P, RajBhandary, U L

We recently described mutants of Escherichia coli initiator tRNA that suppress amber termination codons (UAG) in E. coli. These mutants have changes in the anticodon sequence (CAU----CUA) that allow...

Initiation of protein synthesis from a termination codon.

Varshney, U, RajBhandary, U L

We show that the amber termination codon UAG can initiate protein synthesis in Escherichia coli. We mutated the initiation codon AUG of the chloramphenicol acetyltransferase (CAT) gene to UAG...

Structural studies on tRNA acceptor stem microhelices: exchange of the discriminator base A73 for G in human tRNALeu switches the acceptor specificity from leucine to serine possibly by decreasing the stability of the terminal G1-C72 base pair.

Metzger, A U, Heckl, M, Willbold, D, Breitschopf, K, RajBhandary, U L, Rösch, P, ...

Correct recognition of transfer RNAs (tRNAs) by aminoacyl-tRNA synthetases (aaRS) is crucial to the maintenance of translational fidelity. The discriminator base A73 in human tRNALeuis critical for...

An anticodon sequence mutant of Escherichia coli initiator tRNA: possible importance of a newly acquired base modification next to the anticodon on its activity in initiation.

Mangroo, D, Limbach, P A, McCloskey, J A, RajBhandary, U L

Initiator tRNAs from eubacteria and chloroplasts lack a base modification next to the anticodon. This is in contrast to virtually all other tRNAs from these sources. We show that a mutant Escherichia...

Developmental regulation of the gene for formate dehydrogenase in Neurospora crassa.

Chow, C M, RajBhandary, U L

We have isolated and characterized a gene, fdh, from Neurospora crassa which is developmentally regulated and which produces formate dehydrogenase activity when expressed in Escherichia coli. The...

Role of methionine and formylation of initiator tRNA in initiation of protein synthesis in Escherichia coli.

Varshney, U, RajBhandary, U L

We showed recently that a mutant of Escherichia coli initiator tRNA with a CAU-->CUA anticodon sequence change can initiate protein synthesis from UAG by using formylglutamine instead of...

Escherichia coli B lacks one of the two initiator tRNA species present in E. coli K-12.

Mandal, N, RajBhandary, U L

We show that the metY locus which specifies tRNA(2fMet) in Escherichia coli K-12 specifies tRNA(1fMet) in E. coli B. This conclusion is based on results of Southern blot analysis of E. coli B and...

Amber suppression in mammalian cells dependent upon expression of an Escherichia coli aminoacyl-tRNA synthetase gene.

Drabkin, H J, Park, H J, RajBhandary, U L

As an approach to inducible suppression of nonsense mutations in mammalian and in higher eukaryotic cells, we have analyzed the expression of an Escherichia coli glutamine-inserting amber suppressor...

The A1 x U72 base pair conserved in eukaryotic initiator tRNAs is important specifically for binding to the eukaryotic translation initiation factor eIF2.

Farruggio, D, Chaudhuri, J, Maitra, U, RajBhandary, U L

The formation of a specific ternary complex between eukaryotic initiation factor 2 (eIF2), the initiator methionyl-tRNA (Met-tRNA), and GTP is a critical step in translation initiation in the...

Functional suppression in mammalian cells of nonsense mutations in the herpes simplex virus thymidine kinase gene by suppressor tRNA genes.

Summers, W P, Summers, W C, Laski, F A, RajBhandary, U L, Sharp, P A

A nonsense mutation (UAG) in the thymidine kinase gene of herpes simplex virus type 1 can be suppressed in vivo to produce active thymidine kinase by prior infection with a defective simian virus 40...

Construction, stable transformation, and function of an amber suppressor tRNA gene in Drosophila melanogaster.

Laski, F A, Ganguly, S, Sharp, P A, RajBhandary, U L, Rubin, G M

Drosophila melanogaster strains with a stably incorporated amber suppressor tRNA gene have been generated. A tRNATyr gene was site specifically mutated to produce an anticodon sequence that...

Mutants of Escherichia coli formylmethionine tRNA: a single base change enables initiator tRNA to act as an elongator in vitro.

Seong, B L, RajBhandary, U L

We show that the absence of a Watson-Crick base pair at the end of the amino acid acceptor stem, which is a hallmark of all prokaryotic initiator tRNAs, is one of the key features that prevents them...

Escherichia coli formylmethionine tRNA: mutations in GGGCCC sequence conserved in anticodon stem of initiator tRNAs affect initiation of protein synthesis and conformation of anticodon loop.

Seong, B L, RajBhandary, U L

We have generated mutants of Escherichia coli formylmethionine initiator tRNA in which one, two, and all three G X C base pairs in the GGGCCC sequence in the anticodon stem are changed to those found...

Bacteriorhodopsin: partial sequence of mRNA provides amino acid sequence in the precursor region.

Chang, S H, Majumdar, A, Dunn, R, Makabe, O, RajBhandary, U L, Khorana, H G, ...

mRNA for bacteriorhodopsin from Halobacterium halobium has been partially purified. By using this mRNA as template in the presence of reverse transcriptase RNA-dependent DNA nucleotidyltransferase...

Site-specific mutagenesis on cloned DNAs: generation of a mutant of Escherichia coli tyrosine suppressor tRNA in which the sequence G-T-T-C corresponding to the universal G-T-pseudouracil-C sequence of tRNAs is changed to G-A-T-C.

Kudo, I, Leineweber, M, RajBhandary, U L

We have cloned the Escherichia coli tyrosine-inserting amber suppressor tRNA gene into the recombinant single-strand phage M12mp3. By using the M13mp3SuIII+ recombinant phage DNA as template and an...

Nucleotide sequence of wheat germ cytoplasmic initiator methionine transfer ribonucleic acid.

Ghosh, H P, Ghosh, K, Simsek, M, RajBhandary, U L

The primary sequence of wheat germ initiator tRNA has been determined using in vitro labelling techniques. The sequence is: pAUCAGAGUm1Gm2GCGCAG CGGAAGCGUm2GG psi GGGCCCAUt6AACCCACAGm7GDm5Cm5CCAGGA...

Expression of a X. laevis tRNATyr gene in mammalian cells.

Laski, F A, Alzner-DeWeerd, B, RajBhandary, U L, Sharp, P A

Expression of a X. laevis tRNATyr gene has been studied in mammalian cells. This tRNATyr gene has a 13 base intervening sequence adjacent to its anticodon. A fragment containing the tRNATyr gene was...

The nucleotide sequence of phenylalanine tRNA from the cytoplasm of Neurospora crassa.

Alzner-DeWeerd, B, Hecker, L I, Barnett, W E, RajBhandary, U L

The phenylalanine tRNA from the cytoplasm of Neurospora crassa has been purified and sequenced. The sequence is: pGCGGGUUUAm2GCUCA (N)...

Chemical reactivity of E. coli 5S RNA in situ in the 50S ribosomal subunit.

Silberklang, M, RajBhandary, U L, Lück, A, Erdmann, V A

E. coli 50S ribosomal subunits were reacted with monoperphthalic acid under conditions in which non-base paired adenines are modified to their 1-N-oxides. 5S RNA was isolated from such chemically...

The nucleotide sequence of blue-green algae phenylalanine-tRNA and the evolutionary origin of chloroplasts.

Hecker, L I, Barnett, W E, Lin, F K, Furr, T D, Heckman, J E, RajBhandary, U L, ...

Phenylalanine tRNA from the blue-green alga, Agmenellum quadruplicatum, has been purified to homogeneity. The nucleotide sequence of this tRNA was determined to be: (see tests) Comparisons of the...

The nucleotide sequence of Euglena cytoplasmic phenylalanine transfer RNA. Evidence for possible classifications of Euglena among the animal rather than the plant kingdom.

Chang, S H, Hecker, L I, Brum, C K, Schnabel, J J, Heckman, J E, Silberklang, M, ...

The nucleotide sequence of cytoplasmic phenylalanine tRNA from Euglena gracilis has been elucidated using procedures described previously for the corresponding chloroplastic tRNA [Cell, 9, 717...

The nucleotide sequence of the initiator tRNA from Drosophila melanogaster.

Silverman, S, Heckman, J, Cowling, G J, Delaney, A D, Dunn, R J, Gillam, I C, ...

The nucleotide sequence of Drosophila melanogaster methionine tRNAi was determined to be:...

Sequence analysis of 5'[32P] labeled mRNA and tRNA using polyacrylamide gel electrophoresis.

Lockard, R E, Alzner-Deweerd, B, Heckman, J E, MacGee, J, Tabor, M W, RajBhandary, U L

Sequence analysis of 5'-[32P] labeled tRNA and eukaryotic mRNA using an adaptation of a method recently described by Donis-Keller, Maxam and Gilbert for mapping guanines, adenines and pyrimidines...

The nucleotide sequence of formylmethionine tRNA from Mycoplasma mycoides sp. capri.

Walker, R T, RajBhandary, U L

The nucleotide sequence of Mycoplasma mycoides sp. capri PG3 formylmethionine tRNA has been determined, using in vitro labeling techniques, to be pC-G-C-G-G-G-G-s4U-A-G-A-G-C-A-G-U-D...

Formylatable methionine transfer RNA from Mycoplasma: purification and comparison of partial nucleotide sequences with those of other prokaryotic initiator tRNAs.

Walker, R T, RajBhandary, U L

The major species of the formylatable methionine tRNA from Mycoplasma mycoides var capri has been purified. The 5'- and 3'-terminal sequences of the purified tRNA are pC-G- and C-A-A-C-C-AOH,...

The use of nuclease P1 in sequence analysis of end group labeled RNA.

Silberklang, M, Gillum, A M, RajBhandary, U L

A method is described for the direct sequence analysis of 20-25 nucleotides from the termini of 5'- or 3'-end-group [32P] labeled RNA. The method involves partial endonucleolytic digestion of the...

Nucleotide sequence of Neurospora crassa cytoplasmic initiator tRNA.

Gillum, A M, Hecker, L I, Silberklang, M, Schwartzbach, S D, RajBhandary, U L, Barnett, W E

Initiator methionine tRNA from the cytoplasm of Neurospora crassa has been purified and sequenced. The sequence is: pAGCUGCAUm1GGCGCAGCGGAAGCGCM22GCY*GGGCUCAUt6AACCCGGAGm7GU (or D) -...

2′-O-Methyl ribothymidine: a component of rabbit liver lysine transfer RNA

Gross, H.J., Simsek, M., Raba, M., Limburg, K., Heckman, J., RajBhandary, U.L.

One of the lysine transfer RNAs of rabbit liver is shown to contain 2′-O-methyl ribothymidine in place of ribothymidine. This represents the first demonstration of the presence of 2′-O-methyl...

An amber suppressor tRNA gene derived by site-specific mutagenesis: cloning and function in mammalian cells.

Laski, F A, Belagaje, R, RajBhandary, U L, Sharp, P A

We describe the synthesis, cloning, expression, and in vivo function of a suppressor tRNA gene in mammalian cells. By using "primer-directed mutagenesis" on a Xenopus laevis tyrosine tRNA gene cloned...

A transposable element from Halobacterium halobium which inactivates the bacteriorhodopsin gene.

Simsek, M, DasSarma, S, RajBhandary, U L, Khorana, H G

We describe the characterization of a transposable element from an archaebacterium. The bacteriorhodopsin genes from the wild-type and two mutant Halobacterium halobium strains have been cloned as...

The bacteriorhodopsin gene.

Dunn, R, McCoy, J, Simsek, M, Majumdar, A, Chang, S H, Rajbhandary, U L, ...

The bacteriorhodopsin gene has been identified in a 5.3-kilobase restriction endonuclease fragment isolated from Halobacterium halobium DNA, using a cloned cDNA fragment as the probe. Of the 1229...

Expression and function of a human initiator tRNA gene in the yeast Saccharomyces cerevisiae.

Francis, M A, Rajbhandary, U L

We showed previously that the human initiator tRNA gene, in the context of its own 5'- and 3'-flanking sequences, was not expressed in Saccharomyces cerevisiae. Here we show that switching its...

Cytochrome oxidase subunit V gene of Neurospora crassa: DNA sequences, chromosomal mapping, and evidence that the cya-4 locus specifies the structural gene for subunit V.

Sachs, M S, Bertrand, H, Metzenberg, R L, RajBhandary, U L

The sequences of cDNA and genomic DNA clones for Neurospora cytochrome oxidase subunit V show that the protein is synthesized as a 171-amino-acid precursor containing a 27-amino-acid N-terminal...

Saccharomyces cerevisiae SUP53 tRNA gene transcripts are processed by mammalian cell extracts in vitro but are not processed in vivo.

Ganguly, S, Sharp, P A, RajBhandary, U L

We describe the results of our studies of expression of a Saccharomyces cerevisiae amber suppressor tRNA(Leu) gene (SUP53) in mammalian cells in vivo and in cell extracts in vitro. Parallel studies...

Nuclear gene for mitochondrial leucyl-tRNA synthetase of Neurospora crassa: isolation, sequence, chromosomal mapping, and evidence that the leu-5 locus specifies structural information.

Chow, C M, Metzenberg, R L, Rajbhandary, U L

We have isolated and characterized the nuclear gene for the mitochondrial leucyl-tRNA synthetase (LeuRS) of Neurospora crassa and have established that a defect in this structural gene is responsible...

Regulation of the nuclear genes encoding the cytoplasmic and mitochondrial leucyl-tRNA synthetases of Neurospora crassa.

Chow, C M, Rajbhandary, U L

We show that the nuclear genes for the cytoplasmic and mitochondrial leucyl-tRNA synthetase (LeuRS) of Neurospora crassa are distinct in their encoded proteins, codon usage, mRNA levels, and...

Introduction of UAG, UAA, and UGA nonsense mutations at a specific site in the Escherichia coli chloramphenicol acetyltransferase gene: use in measurement of amber, ochre, and opal suppression in mammalian cells.

Capone, J P, Sedivy, J M, Sharp, P A, RajBhandary, U L

We have used oligonucleotide-directed site-specific mutagenesis to convert serine codon 27 of the Escherichia coli chloramphenicol acetyltransferase (cat) gene to UAG, UAA, and UGA nonsense codons....

Mycobacterium tuberculosis Rv2118c codes for a single-component homotetrameric m1A58 tRNA methyltransferase

Varshney, U., Ramesh, V., Madabushi, A., Gaur, R., Subramanya, H. S., RajBhandary, U. L.

Modified nucleosides in tRNAs play important roles in tRNA structure, biosynthesis and function, and serve as crucial determinants of bacterial growth and virulence. In the yeast Saccharomyces...

Interesting and unusual features in the sequence of Neurospora crassa mitochondrial tyrosine transfer RNA.

Heckman, J E, Alzner-Deweerd, B, RajBhandary, U L

The mitochondrial tyrosine tRNA from Neurospora crassa has been sequenced and found to have several interesting features: (i) It resembles prokaryotic rather than eukaryotic tyrosine tRNAs in that it...

High-frequency spontaneous mutation in the bacterio-opsin gene in Halobacterium halobium is mediated by transposable elements.

DasSarma, S, RajBhandary, U L, Khorana, H G

We have recently characterized a transposable element, ISH1, which inactivates the bacterio-opsin (BO) gene in two purple membrane-deficient (Pum-) mutants of Halobacterium halobium. Examination of...

Nucleotide sequence of 5' terminus of alfalfa mosaic virus RNA 4 leading into coat protein cistron.

Koper-Zwarthoff, E C, Lockard, R E, Alzner-deWeerd, B, RajBhandary, U L, Bol, J F

The sequence of the 5'-terminal 74 nucleotides of alfalfa mosaic virus RNA 4, the mRNA for the viral coat protein, has been deduced by using various new techniques for labeling the RNA at the 5' end...

Ribosome-initiator tRNA complex as an intermediate in translation initiation in Escherichia coli revealed by use of mutant initiator tRNAs and specialized ribosomes.

Wu, X Q, Iyengar, P, RajBhandary, U L

For functional studies of mutant Escherichia coli initiator tRNAs in vivo, we previously described a strategy based on the use of tRNA genes carrying an anticodon sequence change from CAU to CUA...

RNA processing in Neurospora crassa mitochondria: use of transfer RNA sequences as signals.

Breitenberger, C A, Browning, K S, Alzner-DeWeerd, B, RajBhandary, U L

We have used RNA gel transfer hybridization, S1 nuclease mapping and primer extension to analyze transcripts derived from several genes in Neurospora crassa mitochondria. The transcripts studied...

Amber, ochre and opal suppressor tRNA genes derived from a human serine tRNA gene.

Capone, J P, Sharp, P A, RajBhandary, U L

Amber, ochre and opal suppressor tRNA genes have been generated by using oligonucleotide directed site-specific mutagenesis to change one or two nucleotides in a human serine tRNA gene. The amber and...

Synthesis of an ochre suppressor tRNA gene and expression in mammalian cells.

Laski, F A, Belagaje, R, Hudziak, R M, Capecchi, M R, Norton, G P, Palese, P, ...

We have used site-specific mutagenesis to change the anticodon of a Xenopus laevis tyrosine tRNA gene so that it would recognize ochre codons. This tRNA gene is expressed when amplified in monkey...

The E35 stopper mutant of Neurospora crassa: precise localization of deletion endpoints in mitochondrial DNA and evidence that the deleted DNA codes for a subunit of NADH dehydrogenase.

De Vries, H, Alzner-DeWeerd, B, Breitenberger, C A, Chang, D D, De Jonge, J C, RajBhandary, U L

Two types of defective mitochondrial DNA molecules with large deletions (5 kbp and 40 kbp) have previously been identified in the stopper mutant, E35, of Neurospora crassa. The junction fragments...

Cytoplasmic Leucyl-Trna Synthetase of Neurospora Crassa Is Not Specified by the Leu-5 Locus

Benarous, R., Chow, C. M., RajBhandary, U. L.

We generated a λgt11 Neurospora crassa cDNA library and screened the library for the cytoplasmic leucyl-tRNA synthetase (cyto LeuRS) clones using cyto LeuRS specific antibody. Two clones, λNCLRSC1...

Role of the three consecutive G:C base pairs conserved in the anticodon stem of initiator tRNAs in initiation of protein synthesis in Escherichia coli.

Mandal, N, Mangroo, D, Dalluge, J J, McCloskey, J A, Rajbhandary, U L

The three consecutive G:C base pairs, G29:C41, G30:C40, and G31:C39, are conserved in the anticodon stem of virtually all initiator tRNAs from eubacteria, eukaryotes, and archaebacteria. We show that...

Intragenic suppression in tRNA: evidence for crosstalk between the D and the T stems.

Ramesh, V, Varshney, U, Rajbhandary, U L

We showed previously that introduction of two of the three unique features of Escherichia coli initiator tRNA onto an elongator methionine tRNA conferred significant activity in initiation....

Altered discrimination of start codons and initiator tRNAs by mutant initiation factor 3.

O'Connor, M, Gregory, S T, Rajbhandary, U L, Dahlberg, A E

IF3 is essential for ensuring the fidelity of the initiation step of translation in bacterial cells. Mutations at residues R99 and R131 in the C-terminal domain of the factor have previously been...