Links between replication and recombination in Saccharomyces cerevisiae: a hypersensitive requirement for homologous recombination in the absence of Rad27 activity - PubMed (original) (raw)
Links between replication and recombination in Saccharomyces cerevisiae: a hypersensitive requirement for homologous recombination in the absence of Rad27 activity
H Debrauwère et al. Proc Natl Acad Sci U S A. 2001.
Abstract
The RAD27 gene of Saccharomyces cerevisiae encodes a 5'-3' flap exo/endonuclease, which plays an important role during DNA replication for Okazaki fragment maturation. Genetic studies have shown that RAD27 is not essential for growth, although rad27 Delta mutants are temperature sensitive. Moreover, they exhibit increased sensitivity to alkylating agents, enhanced spontaneous recombination, and repetitive DNA instability. The conditional lethality conferred by the rad27 Delta mutation indicates that other nuclease(s) can compensate for the absence of Rad27. Indeed, biochemical and genetical analyses indicate that Okazaki fragment processing can be assured by other enzymatic activities or by alternative pathways such as homologous recombination. Here we present the results of a screen that makes use of a synthetic lethality assay to identify functions required for the survival of rad27 Delta strains. Altogether, we confirm that all genes of the Rad52 recombinational repair pathway are required for the survival of rad27 Delta strains at both permissive (23 degrees C) and semipermissive (30 degrees C) temperatures for growth. We also find that several point mutations that confer weaker phenotypes in mitotic than in meiotic cells (rad50S, mre11s) and additional gene deletions (com1/sae2, srs2) exhibit synthetic lethality with rad27 Delta and that rad59 Delta exhibits synergistic effects with rad27 Delta. This and previous studies indicate that homologous recombination is the primary, but not only, pathway that functions to bypass the replication defects that arise in the absence of the Rad27 protein.
Figures
Figure 1
Simplified steps of Okazaki fragment maturation during DNA lagging strand synthesis (for further details see refs. –5). In rad27Δ cells, unprocessed replication intermediates could be resolved either by using an alternative process involving Dna2, Exo1, RNase H(35), and Polδ activities or by the Rad52 recombinational repair pathway.
Similar articles
- The 3'-->5' exonuclease of DNA polymerase delta can substitute for the 5' flap endonuclease Rad27/Fen1 in processing Okazaki fragments and preventing genome instability.
Jin YH, Obert R, Burgers PM, Kunkel TA, Resnick MA, Gordenin DA. Jin YH, et al. Proc Natl Acad Sci U S A. 2001 Apr 24;98(9):5122-7. doi: 10.1073/pnas.091095198. Epub 2001 Apr 17. Proc Natl Acad Sci U S A. 2001. PMID: 11309502 Free PMC article. - Alleles of the homologous recombination gene, RAD59, identify multiple responses to disrupted DNA replication in Saccharomyces cerevisiae.
Liddell LC, Manthey GM, Owens SN, Fu BX, Bailis AM. Liddell LC, et al. BMC Microbiol. 2013 Oct 14;13:229. doi: 10.1186/1471-2180-13-229. BMC Microbiol. 2013. PMID: 24125552 Free PMC article. - Differential suppression of DNA repair deficiencies of Yeast rad50, mre11 and xrs2 mutants by EXO1 and TLC1 (the RNA component of telomerase).
Lewis LK, Karthikeyan G, Westmoreland JW, Resnick MA. Lewis LK, et al. Genetics. 2002 Jan;160(1):49-62. doi: 10.1093/genetics/160.1.49. Genetics. 2002. PMID: 11805044 Free PMC article. - Recombination factors of Saccharomyces cerevisiae.
Sung P, Trujillo KM, Van Komen S. Sung P, et al. Mutat Res. 2000 Jun 30;451(1-2):257-75. doi: 10.1016/s0027-5107(00)00054-3. Mutat Res. 2000. PMID: 10915877 Review. - DNA replication meets genetic exchange: chromosomal damage and its repair by homologous recombination.
Kuzminov A. Kuzminov A. Proc Natl Acad Sci U S A. 2001 Jul 17;98(15):8461-8. doi: 10.1073/pnas.151260698. Proc Natl Acad Sci U S A. 2001. PMID: 11459990 Free PMC article. Review.
Cited by
- Profiling the compendium of changes in Saccharomyces cerevisiae due to mutations that alter availability of the main methyl donor S-Adenosylmethionine.
Remines M, Schoonover MG, Knox Z, Kenwright K, Hoffert KM, Coric A, Mead J, Ampfer J, Seye S, Strome ED. Remines M, et al. G3 (Bethesda). 2024 Apr 3;14(4):jkae002. doi: 10.1093/g3journal/jkae002. G3 (Bethesda). 2024. PMID: 38184845 Free PMC article. - Reliance of Host-Encoded Regulators of Retromobility on Ty1 Promoter Activity or Architecture.
Salinero AC, Emerson S, Cormier TC, Yin J, Morse RH, Curcio MJ. Salinero AC, et al. Front Mol Biosci. 2022 Jul 1;9:896215. doi: 10.3389/fmolb.2022.896215. eCollection 2022. Front Mol Biosci. 2022. PMID: 35847981 Free PMC article. - RNA-cDNA hybrids mediate transposition via different mechanisms.
Todd LA, Hall AC, Pietrobon V, Chan JNY, Laflamme G, Mekhail K. Todd LA, et al. Sci Rep. 2020 Sep 29;10(1):16034. doi: 10.1038/s41598-020-73018-y. Sci Rep. 2020. PMID: 32994470 Free PMC article. - FEN1 endonuclease as a therapeutic target for human cancers with defects in homologous recombination.
Guo E, Ishii Y, Mueller J, Srivatsan A, Gahman T, Putnam CD, Wang JYJ, Kolodner RD. Guo E, et al. Proc Natl Acad Sci U S A. 2020 Aug 11;117(32):19415-19424. doi: 10.1073/pnas.2009237117. Epub 2020 Jul 27. Proc Natl Acad Sci U S A. 2020. PMID: 32719125 Free PMC article. - Gene Expression Noise Produces Cell-to-Cell Heterogeneity in Eukaryotic Homologous Recombination Rate.
Liu J, François JM, Capp JP. Liu J, et al. Front Genet. 2019 May 21;10:475. doi: 10.3389/fgene.2019.00475. eCollection 2019. Front Genet. 2019. PMID: 31164905 Free PMC article.
References
- Jacquier A, Legrain P, Dujon B. Yeast. 1992;8:121–132. - PubMed
- Lieber M R. BioEssays. 1997;19:233–239. - PubMed
- Bambara R A, Murante R S, Henricksen L A. J Biol Chem. 1997;272:4647–4650. - PubMed
- Waga S, Stillman B. Annu Rev Biochem. 1998;67:721–751. - PubMed
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Molecular Biology Databases
Research Materials