Abstract
Saccharomyces cerevisiae RNA polymerase II conditional mutants that selectively disrupt the synthesis of specific mRNAs were isolated. At the permissive temperature, several of the mutants were inositol auxotrophs as a result of inadequate induction of INO1 transcription. The transcriptional defects exhibited by one of these Ino- mutants (rpb2-2) were further investigated. The induction of GAL10 and HIS4 transcription in rpb2-2 strains was similar to that of wild-type strains, in contrast to the lack of induction of INO1 transcription. When shifted to the nonpermissive temperature, cells containing rpb2-2 continued to accumulate some mRNAs but not others. Together, these results indicate that transcription of specific genes can be disrupted by RNA polymerase II mutations. The rpb2-2 allele alters an amino acid residue that occurs in a highly conserved segment of the RPB2 protein and that is shared by homologous subunits in other species.
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- 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]
- Arndt K. T., Styles C. A., Fink G. R. A suppressor of a HIS4 transcriptional defect encodes a protein with homology to the catalytic subunit of protein phosphatases. Cell. 1989 Feb 24;56(4):527–537. doi: 10.1016/0092-8674(89)90576-x. [DOI] [PubMed] [Google Scholar]
- Barany F. Two-codon insertion mutagenesis of plasmid genes by using single-stranded hexameric oligonucleotides. Proc Natl Acad Sci U S A. 1985 Jun;82(12):4202–4206. doi: 10.1073/pnas.82.12.4202. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Carle G. F., Olson M. V. An electrophoretic karyotype for yeast. Proc Natl Acad Sci U S A. 1985 Jun;82(11):3756–3760. doi: 10.1073/pnas.82.11.3756. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Coulter D. E., Greenleaf A. L. Properties of mutationally altered RNA polymerases II of Drosophila. J Biol Chem. 1982 Feb 25;257(4):1945–1952. [PubMed] [Google Scholar]
- Falkenburg D., Dworniczak B., Faust D. M., Bautz E. K. RNA polymerase II of Drosophila. Relation of its 140,000 Mr subunit to the beta subunit of Escherichia coli RNA polymerase. J Mol Biol. 1987 Jun 20;195(4):929–937. doi: 10.1016/0022-2836(87)90496-7. [DOI] [PubMed] [Google Scholar]
- Glass R. E., Honda A., Ishihama A. Genetic studies on the beta subunit of Escherichia coli RNA polymerase. IX. The role of the carboxy-terminus in enzyme assembly. Mol Gen Genet. 1986 Jun;203(3):492–495. doi: 10.1007/BF00422075. [DOI] [PubMed] [Google Scholar]
- Glass R. E., Jones S. T., Nene V., Nomura T., Fujita N., Ishihama A. Genetic studies on the beta subunit of Escherichia coli RNA polymerase. VIII. Localisation of a region involved in promoter selectivity. Mol Gen Genet. 1986 Jun;203(3):487–491. doi: 10.1007/BF00422074. [DOI] [PubMed] [Google Scholar]
- Greenleaf A. L. Amanitin-resistant RNA polymerase II mutations are in the enzyme's largest subunit. J Biol Chem. 1983 Nov 25;258(22):13403–13406. [PubMed] [Google Scholar]
- Greenleaf A. L., Weeks J. R., Voelker R. A., Ohnishi S., Dickson B. Genetic and biochemical characterization of mutants at an RNA polymerase II locus in D. melanogaster. Cell. 1980 Oct;21(3):785–792. doi: 10.1016/0092-8674(80)90441-9. [DOI] [PubMed] [Google Scholar]
- Guarente L. Regulatory proteins in yeast. Annu Rev Genet. 1987;21:425–452. doi: 10.1146/annurev.ge.21.120187.002233. [DOI] [PubMed] [Google Scholar]
- Guarente L. UASs and enhancers: common mechanism of transcriptional activation in yeast and mammals. Cell. 1988 Feb 12;52(3):303–305. doi: 10.1016/s0092-8674(88)80020-5. [DOI] [PubMed] [Google Scholar]
- Hirsch J. P., Henry S. A. Expression of the Saccharomyces cerevisiae inositol-1-phosphate synthase (INO1) gene is regulated by factors that affect phospholipid synthesis. Mol Cell Biol. 1986 Oct;6(10):3320–3328. doi: 10.1128/mcb.6.10.3320. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hoffman C. S., Winston F. A ten-minute DNA preparation from yeast efficiently releases autonomous plasmids for transformation of Escherichia coli. Gene. 1987;57(2-3):267–272. doi: 10.1016/0378-1119(87)90131-4. [DOI] [PubMed] [Google Scholar]
- Ingles C. J., Himmelfarb H. J., Shales M., Greenleaf A. L., Friesen J. D. Identification, molecular cloning, and mutagenesis of Saccharomyces cerevisiae RNA polymerase genes. Proc Natl Acad Sci U S A. 1984 Apr;81(7):2157–2161. doi: 10.1073/pnas.81.7.2157. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Ma H., Kunes S., Schatz P. J., Botstein D. Plasmid construction by homologous recombination in yeast. Gene. 1987;58(2-3):201–216. doi: 10.1016/0378-1119(87)90376-3. [DOI] [PubMed] [Google Scholar]
- 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]
- McKnight S., Tjian R. Transcriptional selectivity of viral genes in mammalian cells. Cell. 1986 Sep 12;46(6):795–805. doi: 10.1016/0092-8674(86)90061-9. [DOI] [PubMed] [Google Scholar]
- Mitchell P. J., Tjian R. Transcriptional regulation in mammalian cells by sequence-specific DNA binding proteins. Science. 1989 Jul 28;245(4916):371–378. doi: 10.1126/science.2667136. [DOI] [PubMed] [Google Scholar]
- Mortin M. A., Kaufman T. C. Developmental effects of a temperature-sensitive RNA polymerase II mutation in Drosophila melanogaster. Dev Biol. 1984 Jun;103(2):343–354. doi: 10.1016/0012-1606(84)90323-3. [DOI] [PubMed] [Google Scholar]
- Mortin M. A., Kaufman T. C. Developmental genetics of a temperature-sensitive RNA polymerase II mutation in Drosophila melanogaster. Mol Gen Genet. 1982;187(1):120–125. doi: 10.1007/BF00384394. [DOI] [PubMed] [Google Scholar]
- Mortin M. A., Lefevre G., Jr An RNA polymerase II mutation in Drosophila melanogaster that mimics ultrabithorax. Chromosoma. 1981;82(2):237–247. doi: 10.1007/BF00286108. [DOI] [PubMed] [Google Scholar]
- Nomura T., Ishihama A., Kajitani M., Takahashi T., Nakada N., Yoshinaga K. Promoter selectivity of Escherichia coli RNA polymerase. II: Altered promoter selection by mutant holoenzymes. Mol Gen Genet. 1984;193(1):8–16. doi: 10.1007/BF00327407. [DOI] [PubMed] [Google Scholar]
- Nonet M., Scafe C., Sexton J., Young R. Eucaryotic RNA polymerase conditional mutant that rapidly ceases mRNA synthesis. Mol Cell Biol. 1987 May;7(5):1602–1611. doi: 10.1128/mcb.7.5.1602. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ovchinnikov Y. A., Monastyrskaya G. S., Gubanov V. V., Guryev S. O., Chertov OYu, Modyanov N. N., Grinkevich V. A., Makarova I. A., Marchenko T. V., Polovnikova I. N. The primary structure of Escherichia coli RNA polymerase. Nucleotide sequence of the rpoB gene and amino-acid sequence of the beta-subunit. Eur J Biochem. 1981 Jun 1;116(3):621–629. doi: 10.1111/j.1432-1033.1981.tb05381.x. [DOI] [PubMed] [Google Scholar]
- 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]
- Rogalski T. M., Riddle D. L. A Caenorhabditis elegans RNA polymerase II gene, ama-1 IV, and nearby essential genes. Genetics. 1988 Jan;118(1):61–74. doi: 10.1093/genetics/118.1.61. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Sentenac A. Eukaryotic RNA polymerases. CRC Crit Rev Biochem. 1985;18(1):31–90. doi: 10.3109/10409238509082539. [DOI] [PubMed] [Google Scholar]
- Simpson R. B. The molecular topography of RNA polymerase-promoter interaction. Cell. 1979 Oct;18(2):277–285. doi: 10.1016/0092-8674(79)90047-3. [DOI] [PubMed] [Google Scholar]
- St John T. P., Davis R. W. The organization and transcription of the galactose gene cluster of Saccharomyces. J Mol Biol. 1981 Oct 25;152(2):285–315. doi: 10.1016/0022-2836(81)90244-8. [DOI] [PubMed] [Google Scholar]
- Struhl K., Hill D. E. Two related regulatory sequences are required for maximal induction of Saccharomyces cerevisiae his3 transcription. Mol Cell Biol. 1987 Jan;7(1):104–110. doi: 10.1128/mcb.7.1.104. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- 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]
- 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]
- Yura T., Ishihama A. Genetics of bacterial RNA polymerases. Annu Rev Genet. 1979;13:59–97. doi: 10.1146/annurev.ge.13.120179.000423. [DOI] [PubMed] [Google Scholar]