Preferential repair of cyclobutane pyrimidine dimers in the transcribed strand of a gene in yeast chromosomes and plasmids is dependent on transcription.
PDF (1.1M)
Journal: 1992/December - Proceedings of the National Academy of Sciences of the United States of America
ISSN: 0027-8424
PUBMED: 1438266
Abstract:
While preferential repair of the transcribed strands within active genes has been demonstrated in organisms as diverse as humans and Escherichia coli, it has not previously been shown to occur in chromosomal genes in the yeast Saccharomyces cerevisiae. We found that repair of cyclobutane pyrimidine dimers in the transcribed strand of the expressed RPB2 gene in the chromosome of a repair-proficient strain is much more rapid than that in the nontranscribed strand. Furthermore, a copy of the RPB2 gene borne on a centromeric ARS1 plasmid showed the same strand bias in repair. To investigate the relation of this strand bias to transcription, we studied repair in a yeast strain with the temperature-sensitive mutation, rpb1-1, in the largest subunit of RNA polymerase II. When exponentially growing rpb1-1 cells are shifted to the nonpermissive temperature, they rapidly cease mRNA synthesis. At the permissive temperature, both rpb1-1 and the wild-type, parental cells exhibited rapid, proficient repair in the transcribed strand of chromosomal and plasmid-borne copies of the RPB2 gene. At the nonpermissive temperature, the rate of repair in the transcribed strand in rpb1-1 cells was reduced to that in the nontranscribed strand. These findings establish the dependence of strand bias in repair on transcription by RNA polymerase II in the chromosomes and in plasmids, and they validate the use of plasmids for analysis of the relation of repair to transcription in yeast.
Open in
PUBMED | PMC | Google Scholar | Wikipedia
Relations:
Content
Citations
(72)
References
(29)
Chemicals
(3)
Organisms
(1)
Processes
(6)
Anatomy
(2)
Affiliates
(1)
Similar articles
Articles by the same authors
Discussion board
Proc Natl Acad Sci U S A 89(22): 10696-10700
PMID: 1438266

Preferential repair of cyclobutane pyrimidine dimers in the transcribed strand of a gene in yeast chromosomes and plasmids is dependent on transcription.

Abstract

While preferential repair of the transcribed strands within active genes has been demonstrated in organisms as diverse as humans and Escherichia coli, it has not previously been shown to occur in chromosomal genes in the yeast Saccharomyces cerevisiae. We found that repair of cyclobutane pyrimidine dimers in the transcribed strand of the expressed RPB2 gene in the chromosome of a repair-proficient strain is much more rapid than that in the nontranscribed strand. Furthermore, a copy of the RPB2 gene borne on a centromeric ARS1 plasmid showed the same strand bias in repair. To investigate the relation of this strand bias to transcription, we studied repair in a yeast strain with the temperature-sensitive mutation, rpb1-1, in the largest subunit of RNA polymerase II. When exponentially growing rpb1-1 cells are shifted to the nonpermissive temperature, they rapidly cease mRNA synthesis. At the permissive temperature, both rpb1-1 and the wild-type, parental cells exhibited rapid, proficient repair in the transcribed strand of chromosomal and plasmid-borne copies of the RPB2 gene. At the nonpermissive temperature, the rate of repair in the transcribed strand in rpb1-1 cells was reduced to that in the nontranscribed strand. These findings establish the dependence of strand bias in repair on transcription by RNA polymerase II in the chromosomes and in plasmids, and they validate the use of plasmids for analysis of the relation of repair to transcription in yeast.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (1.1M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.
icon of scanned page 10696
10696
icon of scanned page 10697
10697
icon of scanned page 10698
10698
icon of scanned page 10699
10699
icon of scanned page 10700
10700

Images in this article

Image
Image
on p.10697
Image
Image
on p.10698
Click on the image to see a larger version.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Terleth C, van de Putte P, Brouwer J. New insights in DNA repair: preferential repair of transcriptionally active DNA. Mutagenesis. 1991 Mar;6(2):103–111. [PubMed] [Google Scholar]
  • Mellon I, Spivak G, Hanawalt PC. Selective removal of transcription-blocking DNA damage from the transcribed strand of the mammalian DHFR gene. Cell. 1987 Oct 23;51(2):241–249. [PubMed] [Google Scholar]
  • Mellon I, Hanawalt PC. Induction of the Escherichia coli lactose operon selectively increases repair of its transcribed DNA strand. Nature. 1989 Nov 2;342(6245):95–98. [PubMed] [Google Scholar]
  • Selby CP, Sancar A. Gene- and strand-specific repair in vitro: partial purification of a transcription-repair coupling factor. Proc Natl Acad Sci U S A. 1991 Sep 15;88(18):8232–8236.[PMC free article] [PubMed] [Google Scholar]
  • Terleth C, van Sluis CA, van de Putte P. Differential repair of UV damage in Saccharomyces cerevisiae. Nucleic Acids Res. 1989 Jun 26;17(12):4433–4439.[PMC free article] [PubMed] [Google Scholar]
  • Terleth C, Schenk P, Poot R, Brouwer J, van de Putte P. Differential repair of UV damage in rad mutants of Saccharomyces cerevisiae: a possible function of G2 arrest upon UV irradiation. Mol Cell Biol. 1990 Sep;10(9):4678–4684.[PMC free article] [PubMed] [Google Scholar]
  • Terleth C, Waters R, Brouwer J, van de Putte P. Differential repair of UV damage in Saccharomyces cerevisiae is cell cycle dependent. EMBO J. 1990 Sep;9(9):2899–2904.[PMC free article] [PubMed] [Google Scholar]Retracted
  • Smerdon MJ, Thoma F. Site-specific DNA repair at the nucleosome level in a yeast minichromosome. Cell. 1990 May 18;61(4):675–684. [PubMed] [Google Scholar]
  • Smerdon MJ, Bedoyan J, Thoma F. DNA repair in a small yeast plasmid folded into chromatin. Nucleic Acids Res. 1990 Apr 25;18(8):2045–2051.[PMC free article] [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.[PMC free article] [PubMed] [Google Scholar]
  • Sweetser D, Nonet M, Young RA. Prokaryotic and eukaryotic RNA polymerases have homologous core subunits. Proc Natl Acad Sci U S A. 1987 Mar;84(5):1192–1196.[PMC free article] [PubMed] [Google Scholar]
  • 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.[PMC free article] [PubMed] [Google Scholar]
  • Kuo CL, Campbell JL. Cloning of Saccharomyces cerevisiae DNA replication genes: isolation of the CDC8 gene and two genes that compensate for the cdc8-1 mutation. Mol Cell Biol. 1983 Oct;3(10):1730–1737.[PMC free article] [PubMed] [Google Scholar]
  • Bohr VA, Smith CA, Okumoto DS, Hanawalt PC. DNA repair in an active gene: removal of pyrimidine dimers from the DHFR gene of CHO cells is much more efficient than in the genome overall. Cell. 1985 Feb;40(2):359–369. [PubMed] [Google Scholar]
  • Christians FC, Hanawalt PC. Inhibition of transcription and strand-specific DNA repair by alpha-amanitin in Chinese hamster ovary cells. Mutat Res. 1992 Aug;274(2):93–101. [PubMed] [Google Scholar]
  • Leadon SA, Lawrence DA. Preferential repair of DNA damage on the transcribed strand of the human metallothionein genes requires RNA polymerase II. Mutat Res. 1991 Jul;255(1):67–78. [PubMed] [Google Scholar]
  • Carreau M, Hunting D. Transcription-dependent and independent DNA excision repair pathways in human cells. Mutat Res. 1992 Jun;274(1):57–64. [PubMed] [Google Scholar]
  • Marczynski GT, Jaehning JA. A transcription map of a yeast centromere plasmid: unexpected transcripts and altered gene expression. Nucleic Acids Res. 1985 Dec 9;13(23):8487–8506.[PMC free article] [PubMed] [Google Scholar]
  • Nasmyth KA. The regulation of yeast mating-type chromatin structure by SIR: an action at a distance affecting both transcription and transposition. Cell. 1982 Sep;30(2):567–578. [PubMed] [Google Scholar]
  • Selby CP, Witkin EM, Sancar A. Escherichia coli mfd mutant deficient in "mutation frequency decline" lacks strand-specific repair: in vitro complementation with purified coupling factor. Proc Natl Acad Sci U S A. 1991 Dec 15;88(24):11574–11578.[PMC free article] [PubMed] [Google Scholar]
  • Woychik NA, Liao SM, Kolodziej PA, Young RA. Subunits shared by eukaryotic nuclear RNA polymerases. Genes Dev. 1990 Mar;4(3):313–323. [PubMed] [Google Scholar]
  • Armstrong JD, Kunz BA. Site and strand specificity of UVB mutagenesis in the SUP4-o gene of yeast. Proc Natl Acad Sci U S A. 1990 Nov;87(22):9005–9009.[PMC free article] [PubMed] [Google Scholar]
  • Clark RF, Cho KW, Weinmann R, Hamkalo BA. Preferential distribution of active RNA polymerase II molecules in the nuclear periphery. Gene Expr. 1991 Apr;1(1):61–70.[PMC free article] [PubMed] [Google Scholar]
  • Witkin EM. Radiation-induced mutations and their repair. Science. 1966 Jun 3;152(3727):1345–1353. [PubMed] [Google Scholar]
  • Matson SW. Escherichia coli DNA helicase II (uvrD gene product) catalyzes the unwinding of DNA.RNA hybrids in vitro. Proc Natl Acad Sci U S A. 1989 Jun;86(12):4430–4434.[PMC free article] [PubMed] [Google Scholar]
  • Weeda G, van Ham RC, Masurel R, Westerveld A, Odijk H, de Wit J, Bootsma D, van der Eb AJ, Hoeijmakers JH. Molecular cloning and biological characterization of the human excision repair gene ERCC-3. Mol Cell Biol. 1990 Jun;10(6):2570–2581.[PMC free article] [PubMed] [Google Scholar]
  • Bailly V, Sung P, Prakash L, Prakash S. DNA.RNA helicase activity of RAD3 protein of Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1991 Nov 1;88(21):9712–9716.[PMC free article] [PubMed] [Google Scholar]
  • Gulyas KD, Donahue TF. SSL2, a suppressor of a stem-loop mutation in the HIS4 leader encodes the yeast homolog of human ERCC-3. Cell. 1992 Jun 12;69(6):1031–1042. [PubMed] [Google Scholar]
  • Venema J, Mullenders LH, Natarajan AT, van Zeeland AA, Mayne LV. The genetic defect in Cockayne syndrome is associated with a defect in repair of UV-induced DNA damage in transcriptionally active DNA. Proc Natl Acad Sci U S A. 1990 Jun;87(12):4707–4711.[PMC free article] [PubMed] [Google Scholar]
Department of Biological Sciences, Stanford University, CA 94305-5020.
Department of Biological Sciences, Stanford University, CA 94305-5020.
Copyright notice
Abstract
While preferential repair of the transcribed strands within active genes has been demonstrated in organisms as diverse as humans and Escherichia coli, it has not previously been shown to occur in chromosomal genes in the yeast Saccharomyces cerevisiae. We found that repair of cyclobutane pyrimidine dimers in the transcribed strand of the expressed RPB2 gene in the chromosome of a repair-proficient strain is much more rapid than that in the nontranscribed strand. Furthermore, a copy of the RPB2 gene borne on a centromeric ARS1 plasmid showed the same strand bias in repair. To investigate the relation of this strand bias to transcription, we studied repair in a yeast strain with the temperature-sensitive mutation, rpb1-1, in the largest subunit of RNA polymerase II. When exponentially growing rpb1-1 cells are shifted to the nonpermissive temperature, they rapidly cease mRNA synthesis. At the permissive temperature, both rpb1-1 and the wild-type, parental cells exhibited rapid, proficient repair in the transcribed strand of chromosomal and plasmid-borne copies of the RPB2 gene. At the nonpermissive temperature, the rate of repair in the transcribed strand in rpb1-1 cells was reduced to that in the nontranscribed strand. These findings establish the dependence of strand bias in repair on transcription by RNA polymerase II in the chromosomes and in plasmids, and they validate the use of plasmids for analysis of the relation of repair to transcription in yeast.
Collaboration tool especially designed for Life Science professionals.Drag-and-drop any entity to your messages.