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Overexpression of the gene for Rpb7 subunit of yeast RNA polymerase II rescues the phenotypes associated with absence of the largest, nonessential subunit Rpb4

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Abstract

The easily dissociable subcomplex of Rpb4 and Rpb7 subunits of yeast RNA polymerase II has been considered, for long, to play a role in stabilizing Pol II under stress. On the basis of previous genetic and biochemical observations, it was proposed that within the subcomplex one of the functions of Rpb4p is to stabilize the interaction between Rpb7p and the rest of Pol II. We took a direct approach to test the latter possibility by overexpression and mutagenesis ofRPB7 in absence of Rpb4p. We report here the results, which support the latter hypothesis. While it has been previously reported that absence of Rpb4p results in reduction in overall transcription by Pol II, our comparative analysis of RNAs fromRPB4 andrpb4δ cells suggests that there are indeed several genes differentially expressed between the two cells. We propose that the qualitative differences in overall transcription in presence and absence of Rpb4p imply a more active role for Rpb4p in transcription of at least a subset of genes.

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References

  • Ausubel R M., Brent R., Kingston R., Moore D., Seidman, J., Smith J. A. and Struhl K. 1987-1994Current protocols in molecular biology. Wiley, New York.

    Google Scholar 

  • Banuett R 1998 Signaling in the yeasts: an informational cascade with links to the filamentous fungi.Microbiol. Mol. Biol. Rev. 62, 249–274.

    PubMed  CAS  Google Scholar 

  • Choder M. 1993 A growth rate-limiting process in the last growth phase of the yeast life cycle involvesRPB4, a subunit of RNA polymerase II.J. Bacteriol. 175, 6358–6363.

    PubMed  CAS  Google Scholar 

  • Choder M. and Young R. A. 1993 A portion of RNA polymerase II molecules has a component essential for stress response and stress survival.Mol. Cell. Biol. 13, 6984–6991.

    PubMed  CAS  Google Scholar 

  • Rdwards A. M., Kane C. M., Young R. A. and Kornberg R. D. 1991 Two dissociable subunits of yeast RNA polymerase II stimulate the initiation of transcription at a promoterin vitro.j. Biol. Chem. 266, 71–75.

    Google Scholar 

  • Elble R. 1992. A simple and efficient procedure for transformation of yeasts.Biotechniques 13, 18–20.

    PubMed  CAS  Google Scholar 

  • Geitz R. D. and Sugino A. 1988 New yeast-Escherichia coli shuttle vectors constructed within vitro mutagenised yeast genes lacking six base pair restriction sites.Gene 74, 527–534.

    Article  Google Scholar 

  • Gimeno C. J., Ljungdahl P. O., Styles C. A. and Fink G. R. 1993 Characterisation ofSaccharomyces cereviseae pseudohyphal growth. InDimorphic fungi in biology and medicine (ed. H. Vanden Bossche), pp. 83–103. Plenum, New York.

    Google Scholar 

  • Hampsey M. 1998 Molecular genetics of the RNA polymerase II general transcription machinery.Microbiol. Mol. Biol. Rev. 62, 465–503.

    PubMed  CAS  Google Scholar 

  • Ishihama A., Kimuri M. and Motsuzawa H. 1998 Subunits of yeast RNA polymerases: Structure and function.Curr. Opin. Microbiol. 1, 190–196.

    Article  PubMed  CAS  Google Scholar 

  • Khazak V, Sadhale P. P., Woychik N. A., Brent R. and Golemis R. A. 1995 Human RNA polymerase II subunit hsRPB7 functions in yeast and influences stress survival and cell morphology.Mol. Biol. Cell 6, 759–765.

    PubMed  CAS  Google Scholar 

  • Mckune K., Richards K. L., Rdwards A. M., Young R. A. and Woychik N. A. 1993 RPB7, one of two dissociable subunits of yeast RNA polymerase II, is essential for cell viability.Yeast 9, 295–299.

    Article  PubMed  CAS  Google Scholar 

  • Maillet I., Buhler J. M., Sentenac A. and Labarre J. 1999 Rpb4p is necessary for RNA polymerase II activity at high temperature.j. Biol. Chem. 274, 22586–22590.

    Article  PubMed  CAS  Google Scholar 

  • Miyao T. and Woychik N. A. 1998 RNA polymerase subunit RPB5 plays a role in transcriptional activation.Proc. Natl. Acad. Sci. USA 95, 15281–15286.

    Article  PubMed  CAS  Google Scholar 

  • Rosenhech S. and Choder M. 1998 Rpb4, a subunit of RNA polymerase II, enables the enzyme to transcribe at temperature extremesin vitro.J. Bacteriol. 180, 6187–6192.

    Google Scholar 

  • Ruet A., Sentenac A., Fromageot P., Winsor B. and Lacroute R 1980 A mutation of the B220 subunit gene affects the structural and functional properties of yeast RNA polymerase Bin vitro.J. Biol. Chem. 255, 6450–6455.

    PubMed  CAS  Google Scholar 

  • Sadhale P. P. and Woychik N. A. 1994 C25, an essential RNA polymerase III subunit related to the RNA polymerase II subunit RPB7.Mol. Cell. Biol. 14, 6164–6170.

    PubMed  CAS  Google Scholar 

  • Sadhale P. P., Sharma N., Beena P., Katoch A, Acharya N. and Singh S. K. 1998 Modulation of polymerase II composition: a possible mode of transcriptional regulation of stress response in eukaryotes.J. Biosci. 23, 331–335.

    Article  CAS  Google Scholar 

  • Scafe C, Nonet M. and Young R. A. 1990 RNA polymerase II mutants defective in transcription of a subset of genes.Mol. Cell. Biol. 10, 1010–1016.

    PubMed  CAS  Google Scholar 

  • Sheffer A., Varon M. and Choder M. 1999 Rpb7 can interact with RNA polymerase II and support transcription during some stresses independently of Rpb4.Mol. Cell. Biol. 19, 2672–2680.

    PubMed  CAS  Google Scholar 

  • Sherman R, Fink G. R. and Hicks J. B. 1983Methods in yeast genetics. Cold Spring Harbor Laboratory Press, Cold Spring Harbor.

    Google Scholar 

  • Vijayraghavan U., Company M. and Abelson J. 1989 Isolation and characterisation of pre-mRNA splicing mutants ofS. cerevisiae.Genes Dev. 3, 1206–1216.

    Article  PubMed  CAS  Google Scholar 

  • Woychik N. A. and Young R. A. 1989 RNA polymerase II subunit RPB4 is essential for high and low temperature yeast cell growth.Mol. Cell. Biol. 9, 2854–2859.

    PubMed  CAS  Google Scholar 

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Correspondence to Parag P. Sadhale.

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Sharma, N., Sadhale, P.P. Overexpression of the gene for Rpb7 subunit of yeast RNA polymerase II rescues the phenotypes associated with absence of the largest, nonessential subunit Rpb4. J. Genet. 78, 149–156 (1999). https://doi.org/10.1007/BF02934460

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  • DOI: https://doi.org/10.1007/BF02934460

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