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. 1992 Oct;12(10):4314–4326. doi: 10.1128/mcb.12.10.4314

RPC53 encodes a subunit of Saccharomyces cerevisiae RNA polymerase C (III) whose inactivation leads to a predominantly G1 arrest.

C Mann 1, J Y Micouin 1, N Chiannilkulchai 1, I Treich 1, J M Buhler 1, A Sentenac 1
PMCID: PMC360355  PMID: 1406624

Abstract

RPC53 is shown to be an essential gene encoding the C53 subunit specifically associated with yeast RNA polymerase C (III). Temperature-sensitive rpc53 mutants were generated and showed a rapid inhibition of tRNA synthesis after transfer to the restrictive temperature. Unexpectedly, the rpc53 mutants preferentially arrested their cell division in the G1 phase as large, round, unbudded cells. The RPC53 DNA sequence is predicted to code for a hydrophilic M(r)-46,916 protein enriched in charged amino acid residues. The carboxy-terminal 136 amino acids of C53 are significantly similar (25% identical amino acid residues) to the same region of the human BN51 protein. The BN51 cDNA was originally isolated by its ability to complement a temperature-sensitive hamster cell mutant that undergoes a G1 cell division arrest, as is true for the rpc53 mutants.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Allison L. A., Moyle M., Shales M., Ingles C. J. Extensive homology among the largest subunits of eukaryotic and prokaryotic RNA polymerases. Cell. 1985 Sep;42(2):599–610. doi: 10.1016/0092-8674(85)90117-5. [DOI] [PubMed] [Google Scholar]
  2. Baldari C., Cesareni G. Plasmids pEMBLY: new single-stranded shuttle vectors for the recovery and analysis of yeast DNA sequences. Gene. 1985;35(1-2):27–32. doi: 10.1016/0378-1119(85)90154-4. [DOI] [PubMed] [Google Scholar]
  3. Boeke J. D., LaCroute F., Fink G. R. A positive selection for mutants lacking orotidine-5'-phosphate decarboxylase activity in yeast: 5-fluoro-orotic acid resistance. Mol Gen Genet. 1984;197(2):345–346. doi: 10.1007/BF00330984. [DOI] [PubMed] [Google Scholar]
  4. Carles C., Treich I., Bouet F., Riva M., Sentenac A. Two additional common subunits, ABC10 alpha and ABC10 beta, are shared by yeast RNA polymerases. J Biol Chem. 1991 Dec 15;266(35):24092–24096. [PubMed] [Google Scholar]
  5. Dequard-Chablat M., Riva M., Carles C., Sentenac A. RPC19, the gene for a subunit common to yeast RNA polymerases A (I) and C (III). J Biol Chem. 1991 Aug 15;266(23):15300–15307. [PubMed] [Google Scholar]
  6. Dingwall C., Laskey R. A. Nuclear targeting sequences--a consensus? Trends Biochem Sci. 1991 Dec;16(12):478–481. doi: 10.1016/0968-0004(91)90184-w. [DOI] [PubMed] [Google Scholar]
  7. Elledge S. J., Davis R. W. A family of versatile centromeric vectors designed for use in the sectoring-shuffle mutagenesis assay in Saccharomyces cerevisiae. Gene. 1988 Oct 30;70(2):303–312. doi: 10.1016/0378-1119(88)90202-8. [DOI] [PubMed] [Google Scholar]
  8. Gabrielsen O. S., Sentenac A. RNA polymerase III (C) and its transcription factors. Trends Biochem Sci. 1991 Nov;16(11):412–416. doi: 10.1016/0968-0004(91)90166-s. [DOI] [PubMed] [Google Scholar]
  9. Garcia-Bustos J., Heitman J., Hall M. N. Nuclear protein localization. Biochim Biophys Acta. 1991 Mar 7;1071(1):83–101. doi: 10.1016/0304-4157(91)90013-m. [DOI] [PubMed] [Google Scholar]
  10. Geiduschek E. P., Tocchini-Valentini G. P. Transcription by RNA polymerase III. Annu Rev Biochem. 1988;57:873–914. doi: 10.1146/annurev.bi.57.070188.004301. [DOI] [PubMed] [Google Scholar]
  11. Gill G., Tjian R. A highly conserved domain of TFIID displays species specificity in vivo. Cell. 1991 Apr 19;65(2):333–340. doi: 10.1016/0092-8674(91)90166-v. [DOI] [PubMed] [Google Scholar]
  12. Gudenus R., Mariotte S., Moenne A., Ruet A., Memet S., Buhler J. M., Sentenac A., Thuriaux P. Conditional mutants of RPC160, the gene encoding the largest subunit of RNA polymerase C in Saccharomyces cerevisiae. Genetics. 1988 Jul;119(3):517–526. doi: 10.1093/genetics/119.3.517. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hall M. N., Hereford L., Herskowitz I. Targeting of E. coli beta-galactosidase to the nucleus in yeast. Cell. 1984 Apr;36(4):1057–1065. doi: 10.1016/0092-8674(84)90055-2. [DOI] [PubMed] [Google Scholar]
  14. Hanic-Joyce P. J., Johnston G. C., Singer R. A. Regulated arrest of cell proliferation mediated by yeast prt1 mutations. Exp Cell Res. 1987 Sep;172(1):134–145. doi: 10.1016/0014-4827(87)90100-5. [DOI] [PubMed] [Google Scholar]
  15. Hartwell L. H., Mortimer R. K., Culotti J., Culotti M. Genetic Control of the Cell Division Cycle in Yeast: V. Genetic Analysis of cdc Mutants. Genetics. 1973 Jun;74(2):267–286. doi: 10.1093/genetics/74.2.267. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Huet J., Riva M., Sentenac A., Fromageot P. Yeast RNA polymerase C and its subunits. Specific antibodies as structural and functional probes. J Biol Chem. 1985 Dec 5;260(28):15304–15310. [PubMed] [Google Scholar]
  17. Ito H., Fukuda Y., Murata K., Kimura A. Transformation of intact yeast cells treated with alkali cations. J Bacteriol. 1983 Jan;153(1):163–168. doi: 10.1128/jb.153.1.163-168.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Ittmann M., Greco A., Basilico C. Isolation of the human gene that complements a temperature-sensitive cell cycle mutation in BHK cells. Mol Cell Biol. 1987 Oct;7(10):3386–3393. doi: 10.1128/mcb.7.10.3386. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Jaehning J. A., Woods P. S., Roeder R. G. Purification, properties, and subunit structure of deoxyribonucleic acid-dependent ribonucleic acid polymerase III from uninfected and adenovirus 2-infected KB cells. J Biol Chem. 1977 Dec 10;252(23):8762–8771. [PubMed] [Google Scholar]
  20. James P., Whelen S., Hall B. D. The RET1 gene of yeast encodes the second-largest subunit of RNA polymerase III. Structural analysis of the wild-type and ret1-1 mutant alleles. J Biol Chem. 1991 Mar 25;266(9):5616–5624. [PubMed] [Google Scholar]
  21. Jansen R. P., Hurt E. C., Kern H., Lehtonen H., Carmo-Fonseca M., Lapeyre B., Tollervey D. Evolutionary conservation of the human nucleolar protein fibrillarin and its functional expression in yeast. J Cell Biol. 1991 May;113(4):715–729. doi: 10.1083/jcb.113.4.715. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Johnston G. C., Singer R. A. RNA synthesis and control of cell division in the yeast S. cerevisiae. Cell. 1978 Aug;14(4):951–958. doi: 10.1016/0092-8674(78)90349-5. [DOI] [PubMed] [Google Scholar]
  23. Johnston G. C., Singer R. A. Ribosomal precursor RNA metabolism and cell division in the yeast Saccharomyces cerevisiae. Mol Gen Genet. 1980;178(2):357–360. doi: 10.1007/BF00270484. [DOI] [PubMed] [Google Scholar]
  24. Kolodziej P. A., Young R. A. Mutations in the three largest subunits of yeast RNA polymerase II that affect enzyme assembly. Mol Cell Biol. 1991 Sep;11(9):4669–4678. doi: 10.1128/mcb.11.9.4669. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Kolodziej P., Young R. A. RNA polymerase II subunit RPB3 is an essential component of the mRNA transcription apparatus. Mol Cell Biol. 1989 Dec;9(12):5387–5394. doi: 10.1128/mcb.9.12.5387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Lee J. Y., Evans C. F., Engelke D. R. Expression of RNase P RNA in Saccharomyces cerevisiae is controlled by an unusual RNA polymerase III promoter. Proc Natl Acad Sci U S A. 1991 Aug 15;88(16):6986–6990. doi: 10.1073/pnas.88.16.6986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Link A. J., Olson M. V. Physical map of the Saccharomyces cerevisiae genome at 110-kilobase resolution. Genetics. 1991 Apr;127(4):681–698. doi: 10.1093/genetics/127.4.681. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Mann C., Buhler J. M., Treich I., Sentenac A. RPC40, a unique gene for a subunit shared between yeast RNA polymerases A and C. Cell. 1987 Feb 27;48(4):627–637. doi: 10.1016/0092-8674(87)90241-8. [DOI] [PubMed] [Google Scholar]
  29. Marck C. 'DNA Strider': a 'C' program for the fast analysis of DNA and protein sequences on the Apple Macintosh family of computers. Nucleic Acids Res. 1988 Mar 11;16(5):1829–1836. doi: 10.1093/nar/16.5.1829. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Moenne A., Camier S., Anderson G., Margottin F., Beggs J., Sentenac A. The U6 gene of Saccharomyces cerevisiae is transcribed by RNA polymerase C (III) in vivo and in vitro. EMBO J. 1990 Jan;9(1):271–277. doi: 10.1002/j.1460-2075.1990.tb08105.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Mosrin C., Riva M., Beltrame M., Cassar E., Sentenac A., Thuriaux P. The RPC31 gene of Saccharomyces cerevisiae encodes a subunit of RNA polymerase C (III) with an acidic tail. Mol Cell Biol. 1990 Sep;10(9):4737–4743. doi: 10.1128/mcb.10.9.4737. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Nasmyth K. A., Reed S. I. Isolation of genes by complementation in yeast: molecular cloning of a cell-cycle gene. Proc Natl Acad Sci U S A. 1980 Apr;77(4):2119–2123. doi: 10.1073/pnas.77.4.2119. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Orr-Weaver T. L., Szostak J. W., Rothstein R. J. Yeast transformation: a model system for the study of recombination. Proc Natl Acad Sci U S A. 1981 Oct;78(10):6354–6358. doi: 10.1073/pnas.78.10.6354. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Pearson W. R., Lipman D. J. Improved tools for biological sequence comparison. Proc Natl Acad Sci U S A. 1988 Apr;85(8):2444–2448. doi: 10.1073/pnas.85.8.2444. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Pearson W. R. Rapid and sensitive sequence comparison with FASTP and FASTA. Methods Enzymol. 1990;183:63–98. doi: 10.1016/0076-6879(90)83007-v. [DOI] [PubMed] [Google Scholar]
  36. Peden K. W., Nathans D. Local mutagenesis within deletion loops of DNA heteroduplexes. Proc Natl Acad Sci U S A. 1982 Dec;79(23):7214–7217. doi: 10.1073/pnas.79.23.7214. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Philippsen P., Thomas M., Kramer R. A., Davis R. W. Unique arrangement of coding sequences for 5 S, 5.8 S, 18 S and 25 S ribosomal RNA in Saccharomyces cerevisiae as determined by R-loop and hybridization analysis. J Mol Biol. 1978 Aug 15;123(3):387–404. doi: 10.1016/0022-2836(78)90086-4. [DOI] [PubMed] [Google Scholar]
  38. Reed S. I. G1-specific cyclins: in search of an S-phase-promoting factor. Trends Genet. 1991 Mar;7(3):95–99. doi: 10.1016/0168-9525(91)90279-Y. [DOI] [PubMed] [Google Scholar]
  39. Riva M., Memet S., Micouin J. Y., Huet J., Treich I., Dassa J., Young R., Buhler J. M., Sentenac A., Fromageot P. Isolation of structural genes for yeast RNA polymerases by immunological screening. Proc Natl Acad Sci U S A. 1986 Mar;83(6):1554–1558. doi: 10.1073/pnas.83.6.1554. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Rose M. D., Novick P., Thomas J. H., Botstein D., Fink G. R. A Saccharomyces cerevisiae genomic plasmid bank based on a centromere-containing shuttle vector. Gene. 1987;60(2-3):237–243. doi: 10.1016/0378-1119(87)90232-0. [DOI] [PubMed] [Google Scholar]
  41. Rothstein R. J. One-step gene disruption in yeast. Methods Enzymol. 1983;101:202–211. doi: 10.1016/0076-6879(83)01015-0. [DOI] [PubMed] [Google Scholar]
  42. Russel M., Kidd S., Kelley M. R. An improved filamentous helper phage for generating single-stranded plasmid DNA. Gene. 1986;45(3):333–338. doi: 10.1016/0378-1119(86)90032-6. [DOI] [PubMed] [Google Scholar]
  43. Sentenac A. Eukaryotic RNA polymerases. CRC Crit Rev Biochem. 1985;18(1):31–90. doi: 10.3109/10409238509082539. [DOI] [PubMed] [Google Scholar]
  44. Sherman F. Getting started with yeast. Methods Enzymol. 1991;194:3–21. doi: 10.1016/0076-6879(91)94004-v. [DOI] [PubMed] [Google Scholar]
  45. Sklar V. E., Roeder R. G. Purification and subunit structure of deoxyribonucleic acid-dependent ribonucleic acid polymerase III from the mouse plasmacytoma, MOPC 315. J Biol Chem. 1976 Feb 25;251(4):1064–1073. [PubMed] [Google Scholar]
  46. Struhl K., Davis R. W. Position effects in Saccharomyces cerevisiae. J Mol Biol. 1981 Nov 5;152(3):569–575. doi: 10.1016/0022-2836(81)90269-2. [DOI] [PubMed] [Google Scholar]
  47. Struhl K. Nucleotide sequence and transcriptional mapping of the yeast pet56-his3-ded1 gene region. Nucleic Acids Res. 1985 Dec 9;13(23):8587–8601. doi: 10.1093/nar/13.23.8587. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Unger M. W., Hartwell L. H. Control of cell division in Saccharomyces cerevisiae by methionyl-tRNA. Proc Natl Acad Sci U S A. 1976 May;73(5):1664–1668. doi: 10.1073/pnas.73.5.1664. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Unger M. W. Methionyl-transfer ribonucleic acid deficiency during G1 arrest of Saccharomyces cerevisiae. J Bacteriol. 1977 Apr;130(1):11–19. doi: 10.1128/jb.130.1.11-19.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Van Ryk D. I., Lee Y., Nazar R. N. Efficient expression and utilization of mutant 5 S rRNA in Saccharomyces cerevisiae. J Biol Chem. 1990 May 25;265(15):8377–8381. [PubMed] [Google Scholar]
  51. Veinot-Drebot L. M., Singer R. A., Johnston G. C. rRNA transcription initiation is decreased by inhibitors of the yeast cell cycle control step "start". J Biol Chem. 1989 Nov 25;264(33):19528–19534. [PubMed] [Google Scholar]
  52. Vignais M. L., Huet J., Buhler J. M., Sentenac A. Contacts between the factor TUF and RPG sequences. J Biol Chem. 1990 Aug 25;265(24):14669–14674. [PubMed] [Google Scholar]
  53. Vollrath D., Davis R. W., Connelly C., Hieter P. Physical mapping of large DNA by chromosome fragmentation. Proc Natl Acad Sci U S A. 1988 Aug;85(16):6027–6031. doi: 10.1073/pnas.85.16.6027. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Werner M., Hermann-Le Denmat S., Treich I., Sentenac A., Thuriaux P. Effect of mutations in a zinc-binding domain of yeast RNA polymerase C (III) on enzyme function and subunit association. Mol Cell Biol. 1992 Mar;12(3):1087–1095. doi: 10.1128/mcb.12.3.1087. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Woolford J. L., Jr Nuclear pre-mRNA splicing in yeast. Yeast. 1989 Nov-Dec;5(6):439–457. doi: 10.1002/yea.320050604. [DOI] [PubMed] [Google Scholar]

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