Skip to main content
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1994 Sep;14(9):6164–6170. doi: 10.1128/mcb.14.9.6164

C25, an essential RNA polymerase III subunit related to the RNA polymerase II subunit RPB7.

P P Sadhale 1, N A Woychik 1
PMCID: PMC359143  PMID: 8065349

Abstract

We identified a partially sequenced Saccharomyces cerevisiae gene which encodes a protein related to the S. cerevisiae RNA polymerase II subunit, RPB7. Several lines of evidence suggest that this related gene, YKL1, encodes the RNA polymerase III subunit C25. C25, like RPB7, is present in submolar ratios, easily dissociates from the enzyme, is essential for cell growth and viability, but is not required in certain transcription assays in vitro. YKL1 has ABF-1 and PAC upstream sequences often present in RNA polymerase subunit genes. The sodium dodecyl sulfate-polyacrylamide gel electrophoresis mobility of the YKL1 gene product is equivalent to that of the RNA polymerase III subunit C25. Finally, a C25 conditional mutant grown at the nonpermissive temperature synthesizes tRNA at reduced rates relative to 5.8S rRNA, a hallmark of all characterized RNA polymerase III mutants.

Full text

PDF

Images in this article

Selected References

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

  1. Abraham P. R., Mulder A., Van 't Riet J., Planta R. J., Raué H. A. Molecular cloning and physical analysis of an 8.2 kb segment of chromosome XI of Saccharomyces cerevisiae reveals five tightly linked genes. Yeast. 1992 Mar;8(3):227–238. doi: 10.1002/yea.320080309. [DOI] [PubMed] [Google Scholar]
  2. 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]
  3. Boeke J. D., Trueheart J., Natsoulis G., Fink G. R. 5-Fluoroorotic acid as a selective agent in yeast molecular genetics. Methods Enzymol. 1987;154:164–175. doi: 10.1016/0076-6879(87)54076-9. [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. Chiannilkulchai N., Stalder R., Riva M., Carles C., Werner M., Sentenac A. RPC82 encodes the highly conserved, third-largest subunit of RNA polymerase C (III) from Saccharomyces cerevisiae. Mol Cell Biol. 1992 Oct;12(10):4433–4440. doi: 10.1128/mcb.12.10.4433. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Della Seta F., Treich I., Buhler J. M., Sentenac A. ABF1 binding sites in yeast RNA polymerase genes. J Biol Chem. 1990 Sep 5;265(25):15168–15175. [PubMed] [Google Scholar]
  7. 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]
  8. Dezélée S., Wyers F., Sentenac A., Fromageot P. Two forms of RNA polymerase B in yeast. Proteolytic conversion in vitro of enzyme BI into BII. Eur J Biochem. 1976 Jun 1;65(2):543–552. doi: 10.1111/j.1432-1033.1976.tb10372.x. [DOI] [PubMed] [Google Scholar]
  9. Edwards A. M., Kane C. M., Young R. A., Kornberg R. D. Two dissociable subunits of yeast RNA polymerase II stimulate the initiation of transcription at a promoter in vitro. J Biol Chem. 1991 Jan 5;266(1):71–75. [PubMed] [Google Scholar]
  10. Gietz R. D., Sugino A. New yeast-Escherichia coli shuttle vectors constructed with in vitro mutagenized yeast genes lacking six-base pair restriction sites. Gene. 1988 Dec 30;74(2):527–534. doi: 10.1016/0378-1119(88)90185-0. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. Ho S. N., Hunt H. D., Horton R. M., Pullen J. K., Pease L. R. Site-directed mutagenesis by overlap extension using the polymerase chain reaction. Gene. 1989 Apr 15;77(1):51–59. doi: 10.1016/0378-1119(89)90358-2. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Klenk H. P., Palm P., Lottspeich F., Zillig W. Component H of the DNA-dependent RNA polymerases of Archaea is homologous to a subunit shared by the three eucaryal nuclear RNA polymerases. Proc Natl Acad Sci U S A. 1992 Jan 1;89(1):407–410. doi: 10.1073/pnas.89.1.407. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. 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]
  16. Leffers H., Gropp F., Lottspeich F., Zillig W., Garrett R. A. Sequence, organization, transcription and evolution of RNA polymerase subunit genes from the archaebacterial extreme halophiles Halobacterium halobium and Halococcus morrhuae. J Mol Biol. 1989 Mar 5;206(1):1–17. doi: 10.1016/0022-2836(89)90519-6. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. Mann C., Micouin J. Y., Chiannilkulchai N., Treich I., Buhler J. M., Sentenac A. RPC53 encodes a subunit of Saccharomyces cerevisiae RNA polymerase C (III) whose inactivation leads to a predominantly G1 arrest. Mol Cell Biol. 1992 Oct;12(10):4314–4326. doi: 10.1128/mcb.12.10.4314. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. McKune K., Woychik N. A. Halobacterial S9 operon contains two genes encoding proteins homologous to subunits shared by eukaryotic RNA polymerases I, II, and III. J Bacteriol. 1994 Aug;176(15):4754–4756. doi: 10.1128/jb.176.15.4754-4756.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. 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]
  21. Muhlrad D., Hunter R., Parker R. A rapid method for localized mutagenesis of yeast genes. Yeast. 1992 Feb;8(2):79–82. doi: 10.1002/yea.320080202. [DOI] [PubMed] [Google Scholar]
  22. Mémet S., Gouy M., Marck C., Sentenac A., Buhler J. M. RPA190, the gene coding for the largest subunit of yeast RNA polymerase A. J Biol Chem. 1988 Feb 25;263(6):2830–2839. [PubMed] [Google Scholar]
  23. Nogi Y., Yano R., Dodd J., Carles C., Nomura M. Gene RRN4 in Saccharomyces cerevisiae encodes the A12.2 subunit of RNA polymerase I and is essential only at high temperatures. Mol Cell Biol. 1993 Jan;13(1):114–122. doi: 10.1128/mcb.13.1.114. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Pühler G., Leffers H., Gropp F., Palm P., Klenk H. P., Lottspeich F., Garrett R. A., Zillig W. Archaebacterial DNA-dependent RNA polymerases testify to the evolution of the eukaryotic nuclear genome. Proc Natl Acad Sci U S A. 1989 Jun;86(12):4569–4573. doi: 10.1073/pnas.86.12.4569. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Ruet A., Sentenac A., Fromageot P., Winsor B., Lacroute F. A mutation of the B220 subunit gene affects the structural and functional properties of yeast RNA polymerase B in vitro. J Biol Chem. 1980 Jul 10;255(13):6450–6455. [PubMed] [Google Scholar]
  26. Scholzen T., Arndt E. The alpha-operon equivalent genome region in the extreme halophilic archaebacterium Haloarcula (Halobacterium) marismortui. J Biol Chem. 1992 Jun 15;267(17):12123–12130. [PubMed] [Google Scholar]
  27. Stettler S., Mariotte S., Riva M., Sentenac A., Thuriaux P. An essential and specific subunit of RNA polymerase III (C) is encoded by gene RPC34 in Saccharomyces cerevisiae. J Biol Chem. 1992 Oct 25;267(30):21390–21395. [PubMed] [Google Scholar]
  28. Sweetser D., Nonet M., Young R. A. Prokaryotic and eukaryotic RNA polymerases have homologous core subunits. Proc Natl Acad Sci U S A. 1987 Mar;84(5):1192–1196. doi: 10.1073/pnas.84.5.1192. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Treich I., Carles C., Riva M., Sentenac A. RPC10 encodes a new mini subunit shared by yeast nuclear RNA polymerases. Gene Expr. 1992;2(1):31–37. [PMC free article] [PubMed] [Google Scholar]
  30. Ulmasov T., Guilfoyle T. J. Sequence of the fifth largest subunit of RNA polymerase II from plants. J Biol Chem. 1992 Nov 15;267(32):23165–23169. [PubMed] [Google Scholar]
  31. Woychik N. A., Lane W. S., Young R. A. Yeast RNA polymerase II subunit RPB9 is essential for growth at temperature extremes. J Biol Chem. 1991 Oct 5;266(28):19053–19055. [PubMed] [Google Scholar]
  32. Woychik N. A., Liao S. M., Kolodziej P. A., Young R. A. Subunits shared by eukaryotic nuclear RNA polymerases. Genes Dev. 1990 Mar;4(3):313–323. doi: 10.1101/gad.4.3.313. [DOI] [PubMed] [Google Scholar]
  33. Woychik N. A., McKune K., Lane W. S., Young R. A. Yeast RNA polymerase II subunit RPB11 is related to a subunit shared by RNA polymerase I and III. Gene Expr. 1993;3(1):77–82. [PMC free article] [PubMed] [Google Scholar]
  34. Woychik N. A., Young R. A. RNA polymerase II subunit RPB10 is essential for yeast cell viability. J Biol Chem. 1990 Oct 15;265(29):17816–17819. [PubMed] [Google Scholar]
  35. Woychik N. A., Young R. A. RNA polymerase II subunit RPB4 is essential for high- and low-temperature yeast cell growth. Mol Cell Biol. 1989 Jul;9(7):2854–2859. doi: 10.1128/mcb.9.7.2854. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Yano R., Nomura M. Suppressor analysis of temperature-sensitive mutations of the largest subunit of RNA polymerase I in Saccharomyces cerevisiae: a suppressor gene encodes the second-largest subunit of RNA polymerase I. Mol Cell Biol. 1991 Feb;11(2):754–764. doi: 10.1128/mcb.11.2.754. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Young R. A. RNA polymerase II. Annu Rev Biochem. 1991;60:689–715. doi: 10.1146/annurev.bi.60.070191.003353. [DOI] [PubMed] [Google Scholar]

Articles from Molecular and Cellular Biology are provided here courtesy of Taylor & Francis

RESOURCES