Abstract
We have screened a Saccharomyces cerevisiae expression library with antibodies against seryl-tRNA synthetase (SerRS) from baker's yeast. In this way we obtained clones which contain serS, the structural gene for seryl-tRNA synthetase. Genomic Southern blots show that the serS gene resides on a 5.0 kb SalI fragment. Nucleotide sequence analysis of the genes revealed a single open reading frame from which we deduced the amino acid sequence of the enzyme consistent with that of two peptides isolated from SerRS. The enzyme is comprised of 462 amino acids consistent with earlier determinations of its molecular weight. The codon usage of serS is typical of abundant yeast proteins. Nuclease S1 analysis of serS mRNA defined the RNA initiation site 20-40 bases downstream from an AT rich sequence containing the TATA box and 21-39 nucleotides upstream of the translation initiation codon. Yeast strains transformed with the cloned gene overproduce seryl-tRNA synthetase in vivo.
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- Bhat T. N., Blow D. M., Brick P., Nyborg J. Tyrosyl-tRNA synthetase forms a mononucleotide-binding fold. J Mol Biol. 1982 Jul 15;158(4):699–709. doi: 10.1016/0022-2836(82)90255-8. [DOI] [PubMed] [Google Scholar]
- Blow D. M., Bhat T. N., Metcalfe A., Risler J. L., Brunie S., Zelwer C. Structural homology in the amino-terminal domains of two aminoacyl-tRNA synthetases. J Mol Biol. 1983 Dec 25;171(4):571–576. doi: 10.1016/0022-2836(83)90044-x. [DOI] [PubMed] [Google Scholar]
- Breton R., Sanfaçon H., Papayannopoulos I., Biemann K., Lapointe J. Glutamyl-tRNA synthetase of Escherichia coli. Isolation and primary structure of the gltX gene and homology with other aminoacyl-tRNA synthetases. J Biol Chem. 1986 Aug 15;261(23):10610–10617. [PubMed] [Google Scholar]
- Broach J. R., Strathern J. N., Hicks J. B. Transformation in yeast: development of a hybrid cloning vector and isolation of the CAN1 gene. Gene. 1979 Dec;8(1):121–133. doi: 10.1016/0378-1119(79)90012-x. [DOI] [PubMed] [Google Scholar]
- Carlson M., Botstein D. Two differentially regulated mRNAs with different 5' ends encode secreted with intracellular forms of yeast invertase. Cell. 1982 Jan;28(1):145–154. doi: 10.1016/0092-8674(82)90384-1. [DOI] [PubMed] [Google Scholar]
- Carter P. J., Winter G., Wilkinson A. J., Fersht A. R. The use of double mutants to detect structural changes in the active site of the tyrosyl-tRNA synthetase (Bacillus stearothermophilus). Cell. 1984 Oct;38(3):835–840. doi: 10.1016/0092-8674(84)90278-2. [DOI] [PubMed] [Google Scholar]
- Chen W., Struhl K. Yeast mRNA initiation sites are determined primarily by specific sequences, not by the distance from the TATA element. EMBO J. 1985 Dec 1;4(12):3273–3280. doi: 10.1002/j.1460-2075.1985.tb04077.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dobson M. J., Tuite M. F., Roberts N. A., Kingsman A. J., Kingsman S. M., Perkins R. E., Conroy S. C., Fothergill L. A. Conservation of high efficiency promoter sequences in Saccharomyces cerevisiae. Nucleic Acids Res. 1982 Apr 24;10(8):2625–2637. doi: 10.1093/nar/10.8.2625. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Erlich H. A., Cohen S. N., McDevitt H. O. Immunological detection and characterization of products translated from cloned DNA fragments. Methods Enzymol. 1979;68:443–453. doi: 10.1016/0076-6879(79)68034-5. [DOI] [PubMed] [Google Scholar]
- Fasiolo F., Bonnet J., Lacroute F. Cloning of the yeast methionyl-tRNA synthetase gene. J Biol Chem. 1981 Mar 10;256(5):2324–2328. [PubMed] [Google Scholar]
- Freedman R., Gibson B., Donovan D., Biemann K., Eisenbeis S., Parker J., Schimmel P. Primary structure of histidine-tRNA synthetase and characterization of hisS transcripts. J Biol Chem. 1985 Aug 25;260(18):10063–10068. [PubMed] [Google Scholar]
- Goto T., Wang J. C. Yeast DNA topoisomerase II is encoded by a single-copy, essential gene. Cell. 1984 Apr;36(4):1073–1080. doi: 10.1016/0092-8674(84)90057-6. [DOI] [PubMed] [Google Scholar]
- Heider H., Gottschalk E., Cramer F. Isolation and characterization of seryl-tRNA synthetase from yeast. Eur J Biochem. 1971 May 11;20(1):144–152. doi: 10.1111/j.1432-1033.1971.tb01372.x. [DOI] [PubMed] [Google Scholar]
- Heyer W. D., Sipiczki M., Kohli J. Replicating plasmids in Schizosaccharomyces pombe: improvement of symmetric segregation by a new genetic element. Mol Cell Biol. 1986 Jan;6(1):80–89. doi: 10.1128/mcb.6.1.80. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hoben P., Royal N., Cheung A., Yamao F., Biemann K., Söll D. Escherichia coli glutaminyl-tRNA synthetase. II. Characterization of the glnS gene product. J Biol Chem. 1982 Oct 10;257(19):11644–11650. [PubMed] [Google Scholar]
- Horwich A. L., Fenton W. A., Williams K. R., Kalousek F., Kraus J. P., Doolittle R. F., Konigsberg W., Rosenberg L. E. Structure and expression of a complementary DNA for the nuclear coded precursor of human mitochondrial ornithine transcarbamylase. Science. 1984 Jun 8;224(4653):1068–1074. doi: 10.1126/science.6372096. [DOI] [PubMed] [Google Scholar]
- Hottinger H., Pearson D., Yamao F., Gamulin V., Cooley L., Cooper T., Söll D. Nonsense suppression in Schizosaccharomyces pombe: the S. pombe Sup3-e tRNASerUGA gene is active in S. cerevisiae. Mol Gen Genet. 1982;188(2):219–224. doi: 10.1007/BF00332678. [DOI] [PubMed] [Google Scholar]
- Ikemura T. Codon usage and tRNA content in unicellular and multicellular organisms. Mol Biol Evol. 1985 Jan;2(1):13–34. doi: 10.1093/oxfordjournals.molbev.a040335. [DOI] [PubMed] [Google Scholar]
- Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
- Lazarides E., Weber K. Actin antibody: the specific visualization of actin filaments in non-muscle cells. Proc Natl Acad Sci U S A. 1974 Jun;71(6):2268–2272. doi: 10.1073/pnas.71.6.2268. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ludmerer S. W., Schimmel P. Cloning of GLN4: an essential gene that encodes glutaminyl-tRNA synthetase in Saccharomyces cerevisiae. J Bacteriol. 1985 Aug;163(2):763–768. doi: 10.1128/jb.163.2.763-768.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Meussdoerffer F., Fink G. R. Structure and expression of two aminoacyl-tRNA synthetase genes from Saccharomyces cerevisiae. J Biol Chem. 1983 May 25;258(10):6293–6299. [PubMed] [Google Scholar]
- Mirande M., Le Corre D., Riva M., Waller J. P. Cloning of yeast lysyl- and phenylalanyl-tRNA synthetase genes. Biochimie. 1986 Jul-Aug;68(7-8):1001–1007. doi: 10.1016/s0300-9084(86)80043-8. [DOI] [PubMed] [Google Scholar]
- Mitchell A. P., Ludmerer S. W. Identification of a glutaminyl-tRNA synthetase mutation Saccharomyces cerevisiae. J Bacteriol. 1984 May;158(2):530–534. doi: 10.1128/jb.158.2.530-534.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Myers A. M., Tzagoloff A. MSW, a yeast gene coding for mitochondrial tryptophanyl-tRNA synthetase. J Biol Chem. 1985 Dec 5;260(28):15371–15377. [PubMed] [Google Scholar]
- Nagata S., Tsunetsugu-Yokota Y., Naito A., Kaziro Y. Molecular cloning and sequence determination of the nuclear gene coding for mitochondrial elongation factor Tu of Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1983 Oct;80(20):6192–6196. doi: 10.1073/pnas.80.20.6192. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nagawa F., Fink G. R. The relationship between the "TATA" sequence and transcription initiation sites at the HIS4 gene of Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1985 Dec;82(24):8557–8561. doi: 10.1073/pnas.82.24.8557. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pape L. K., Koerner T. J., Tzagoloff A. Characterization of a yeast nuclear gene (MST1) coding for the mitochondrial threonyl-tRNA1 synthetase. J Biol Chem. 1985 Dec 5;260(28):15362–15370. [PubMed] [Google Scholar]
- Pape L. K., Tzagoloff A. Cloning and characterization of the gene for the yeast cytoplasmic threonyl-tRNA synthetase. Nucleic Acids Res. 1985 Sep 11;13(17):6171–6183. doi: 10.1093/nar/13.17.6171. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schaack J., Söll D. Transcription of a Drosophila tRNAArg gene in yeast extract: 5'-flanking sequence dependence for transcription in a heterologous system. Nucleic Acids Res. 1985 Apr 25;13(8):2803–2814. doi: 10.1093/nar/13.8.2803. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schatz G., Butow R. A. How are proteins imported into mitochondria? Cell. 1983 Feb;32(2):316–318. doi: 10.1016/0092-8674(83)90450-6. [DOI] [PubMed] [Google Scholar]
- Schimmel P. R., Söll D. Aminoacyl-tRNA synthetases: general features and recognition of transfer RNAs. Annu Rev Biochem. 1979;48:601–648. doi: 10.1146/annurev.bi.48.070179.003125. [DOI] [PubMed] [Google Scholar]
- Sellami M., Fasiolo F., Dirheimer G., Ebel J. P., Gangloff J. Nucleotide sequence of the gene coding for yeast cytoplasmic aspartyl-tRNA synthetase (APS); mapping of the 5' and 3' termini of AspRS mRNA. Nucleic Acids Res. 1986 Feb 25;14(4):1657–1666. doi: 10.1093/nar/14.4.1657. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sellami M., Prévost G., Bonnet J., Dirheimer G., Gangloff J. Isolation and characterization of the yeast aspartyl-tRNA synthetase gene. Gene. 1985;40(2-3):349–352. doi: 10.1016/0378-1119(85)90060-5. [DOI] [PubMed] [Google Scholar]
- Stanley K. K. Solubilization and immune-detection of beta-galactosidase hybrid proteins carrying foreign antigenic determinants. Nucleic Acids Res. 1983 Jun 25;11(12):4077–4092. doi: 10.1093/nar/11.12.4077. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Thonart P., Bechet J., Hilger F., Burny A. Thermosensitive mutations affecting ribonucleic acid polymerases in Saccharomyces cerevisiae. J Bacteriol. 1976 Jan;125(1):25–32. doi: 10.1128/jb.125.1.25-32.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Walter P., Gangloff J., Bonnet J., Boulanger Y., Ebel J. P., Fasiolo F. Primary structure of the Saccharomyces cerevisiae gene for methionyl-tRNA synthetase. Proc Natl Acad Sci U S A. 1983 May;80(9):2437–2441. doi: 10.1073/pnas.80.9.2437. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Weaver R. F., Weissmann C. Mapping of RNA by a modification of the Berk-Sharp procedure: the 5' termini of 15 S beta-globin mRNA precursor and mature 10 s beta-globin mRNA have identical map coordinates. Nucleic Acids Res. 1979 Nov 10;7(5):1175–1193. doi: 10.1093/nar/7.5.1175. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Webster T., Tsai H., Kula M., Mackie G. A., Schimmel P. Specific sequence homology and three-dimensional structure of an aminoacyl transfer RNA synthetase. Science. 1984 Dec 14;226(4680):1315–1317. doi: 10.1126/science.6390679. [DOI] [PubMed] [Google Scholar]
- Willis I., Frendewey D., Nichols M., Hottinger-Werlen A., Schaack J., Söll D. A single base change in the intron of a serine tRNA affects the rate of RNase P cleavage in vitro and suppressor activity in vivo in Saccharomyces cerevisiae. J Biol Chem. 1986 May 5;261(13):5878–5885. [PubMed] [Google Scholar]
- Winter G., Fersht A. R., Wilkinson A. J., Zoller M., Smith M. Redesigning enzyme structure by site-directed mutagenesis: tyrosyl tRNA synthetase and ATP binding. Nature. 1982 Oct 21;299(5885):756–758. doi: 10.1038/299756a0. [DOI] [PubMed] [Google Scholar]
- Young R. A., Davis R. W. Efficient isolation of genes by using antibody probes. Proc Natl Acad Sci U S A. 1983 Mar;80(5):1194–1198. doi: 10.1073/pnas.80.5.1194. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Young R. A., Davis R. W. Yeast RNA polymerase II genes: isolation with antibody probes. Science. 1983 Nov 18;222(4625):778–782. doi: 10.1126/science.6356359. [DOI] [PubMed] [Google Scholar]
- Zelwer C., Risler J. L., Brunie S. Crystal structure of Escherichia coli methionyl-tRNA synthetase at 2.5 A resolution. J Mol Biol. 1982 Feb 15;155(1):63–81. doi: 10.1016/0022-2836(82)90492-2. [DOI] [PubMed] [Google Scholar]



