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. 1993 Feb 11;21(3):469–473. doi: 10.1093/nar/21.3.469

Cloning and sequence determination of the Schizosaccharomyces pombe rpb2 gene encoding the subunit 2 of RNA polymerase II.

M Kawagishi 1, M Yamagishi 1, A Ishihama 1
PMCID: PMC309141  PMID: 8441660

Abstract

The gene, rpb2, encoding the second largest subunit, subunit 2, of RNA polymerase II has been cloned from Schizosaccharomyces pombe using the corresponding gene, RPB2, of Saccharomyces cerevisiae as a probe for cross-hybridization. We have determined the complete nucleotide sequence of rpb2, and parts of the PCR-amplified rpb2 cDNA. The predicted coding sequence of a polypeptide of 1210 amino acid residues with a calculated molecular weight of 138 kilodaltons was interrupted by a short intron. The overall amino acid sequence homology of the S. pombe subunit 2 is 68, 62 and 62% with the corresponding protein from S. cerevisiae, D. melanogaster and H. sapiens, respectively. Southern analysis of the genomic DNA digested with various restriction enzymes showed that rpb2 was present as a single copy in the S. pombe genome. Northern analysis showed that the transcript of rpb2 was about 4 kb in length.

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Selected References

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  1. Acker J., Wintzerith M., Vigneron M., Kédinger C. Primary structure of the second largest subunit of human RNA polymerase II (or B). J Mol Biol. 1992 Aug 20;226(4):1295–1299. doi: 10.1016/0022-2836(92)91071-v. [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. Azuma Y., Yamagishi M., Ueshima R., Ishihama A. Cloning and sequence determination of the Schizosaccharomyces pombe rpb1 gene encoding the largest subunit of RNA polymerase II. Nucleic Acids Res. 1991 Feb 11;19(3):461–468. doi: 10.1093/nar/19.3.461. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Ishihama A. Promoter selectivity of prokaryotic RNA polymerases. Trends Genet. 1988 Oct;4(10):282–286. doi: 10.1016/0168-9525(88)90170-9. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. Kozak M. Comparison of initiation of protein synthesis in procaryotes, eucaryotes, and organelles. Microbiol Rev. 1983 Mar;47(1):1–45. doi: 10.1128/mr.47.1.1-45.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Käufer N. F., Simanis V., Nurse P. Fission yeast Schizosaccharomyces pombe correctly excises a mammalian RNA transcript intervening sequence. Nature. 1985 Nov 7;318(6041):78–80. doi: 10.1038/318078a0. [DOI] [PubMed] [Google Scholar]
  8. Mertins P., Gallwitz D. Nuclear pre-mRNA splicing in the fission yeast Schizosaccharomyces pombe strictly requires an intron-contained, conserved sequence element. EMBO J. 1987 Jun;6(6):1757–1763. doi: 10.1002/j.1460-2075.1987.tb02428.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Nonet M., Sweetser D., Young R. A. Functional redundancy and structural polymorphism in the large subunit of RNA polymerase II. Cell. 1987 Sep 11;50(6):909–915. doi: 10.1016/0092-8674(87)90517-4. [DOI] [PubMed] [Google Scholar]
  10. Padgett R. A., Grabowski P. J., Konarska M. M., Seiler S., Sharp P. A. Splicing of messenger RNA precursors. Annu Rev Biochem. 1986;55:1119–1150. doi: 10.1146/annurev.bi.55.070186.005351. [DOI] [PubMed] [Google Scholar]
  11. Russell P. R. Transcription of the triose-phosphate-isomerase gene of Schizosaccharomyces pombe initiates from a start point different from that in Saccharomyces cerevisiae. Gene. 1985;40(1):125–130. doi: 10.1016/0378-1119(85)90031-9. [DOI] [PubMed] [Google Scholar]
  12. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Scafe C., Martin C., Nonet M., Podos S., Okamura S., Young R. A. Conditional mutations occur predominantly in highly conserved residues of RNA polymerase II subunits. Mol Cell Biol. 1990 Mar;10(3):1270–1275. doi: 10.1128/mcb.10.3.1270. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Sentenac A. Eukaryotic RNA polymerases. CRC Crit Rev Biochem. 1985;18(1):31–90. doi: 10.3109/10409238509082539. [DOI] [PubMed] [Google Scholar]
  15. Shirai T., Go M. RNase-like domain in DNA-directed RNA polymerase II. Proc Natl Acad Sci U S A. 1991 Oct 15;88(20):9056–9060. doi: 10.1073/pnas.88.20.9056. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Sitzler S., Oldenburg I., Petersen G., Bautz E. K. Analysis of the promoter region of the housekeeping gene DmRP140 by sequence comparison of Drosophila melanogaster and Drosophila virilis. Gene. 1991 Apr;100:155–162. doi: 10.1016/0378-1119(91)90361-e. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. 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]
  19. Treich I., Riva M., Sentenac A. Zinc-binding subunits of yeast RNA polymerases. J Biol Chem. 1991 Nov 15;266(32):21971–21976. [PubMed] [Google Scholar]
  20. 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]
  21. 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]
  22. 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]
  23. Woychik N. A., Young R. A. RNA polymerase II: subunit structure and function. Trends Biochem Sci. 1990 Sep;15(9):347–351. doi: 10.1016/0968-0004(90)90074-l. [DOI] [PubMed] [Google Scholar]
  24. 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]

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