ANALYSIS OF MEIOSIS-DEFECTIVE MUTATIONS IN YEAST BY PHYSICAL MONITORING OF RECOMBINATION

Rhona H. Borts ¹, Michael Lichten ¹, and James E. Haber ¹

¹ Rosenstiel Basic Medical Sciences Research Center and Department of Biology, Brandeis University, Waltham, Massachusetts 02254

We have developed a method by which the extent of physical exchange of DNA molecules can be determined throughout meiosis in the yeast Saccharomyces cerevisiae. We have used this technique to analyze the effect of five meiosis-defective mutations (rad6, rad50, rad52, rad57 and spo11 ) on the physical exchange of DNA molecules. In the same experiments, we have also measured other meiotic parameters, such as premeiotic DNA synthesis, commitment to intragenic recombination, haploidization, ascus formation, and viability. rad50 and spo11 diploids make an undetectable amount of physically recombined DNA and <1% of wild-type levels of viable intragenic recombinants. In contrast, diploids homozygous for rad52, rad6 or rad57 all yield significant amounts of novel restriction fragments which arise by recombination. rad57 diploids make nearly wild-type levels of the recombined restriction fragments, although they produce <10% of the wild-type levels of viable intragenic recombinants. rad52 strains are also capable of a significant (33%) amount of exchange of DNA molecules, but make <1% of wild-type levels of viable intragenic recombinants. rad6 diploids are also capable of undergoing a high level of exchange, as measured by the appearance of the recombined restriction fragment. In addition, rad6 diploids show an unusual allele- or locus-specific variability in the level of viable intragenic recombinants produced. Although rad6 diploids produce no viable spores, they are able to complete a significant amount of haploidization upon return to vegetative growth conditions.

This article has been cited by other articles:

				E. Coic, T. Feldman, A. S. Landman, and J. E. Haber Mechanisms of Rad52-Independent Spontaneous and UV-Induced Mitotic Recombination in Saccharomyces cerevisiae Genetics, May 1, 2008; 179(1): 199 - 211. [Abstract] [Full Text] [PDF]

				A. M. Mozlin, C. W. Fung, and L. S. Symington Role of the Saccharomyces cerevisiae Rad51 Paralogs in Sister Chromatid Recombination Genetics, January 1, 2008; 178(1): 113 - 126. [Abstract] [Full Text] [PDF]

				V. Jantsch, L. Tang, P. Pasierbek, A. Penkner, S. Nayak, A. Baudrimont, T. Schedl, A. Gartner, and J. Loidl Caenorhabditis elegans prom-1 Is Required for Meiotic Prophase Progression and Homologous Chromosome Pairing Mol. Biol. Cell, December 1, 2007; 18(12): 4911 - 4920. [Abstract] [Full Text] [PDF]

				E. R. Hoffmann, E. Eriksson, B. J. Herbert, and R. H. Borts MLH1 and MSH2 Promote the Symmetry of Double-Strand Break Repair Events at the HIS4 Hotspot in Saccharomyces cerevisiae Genetics, March 1, 2005; 169(3): 1291 - 1303. [Abstract] [Full Text] [PDF]

				M. Becker, N. Aitcheson, E. Byles, B. Wickstead, E. Louis, and G. Rudenko Isolation of the repertoire of VSG expression site containing telomeres of Trypanosoma brucei 427 using transformation-associated recombination in yeast Genome Res., November 1, 2004; 14(11): 2319 - 2329. [Abstract] [Full Text] [PDF]

				H. Tsubouchi and G. S. Roeder The Budding Yeast Mei5 and Sae3 Proteins Act Together With Dmc1 During Meiotic Recombination Genetics, November 1, 2004; 168(3): 1219 - 1230. [Abstract] [Full Text] [PDF]

				C.-F. Kao, C. Hillyer, T. Tsukuda, K. Henry, S. Berger, and M. A. Osley Rad6 plays a role in transcriptional activation through ubiquitylation of histone H2B Genes & Dev., January 15, 2004; 18(2): 184 - 195. [Abstract] [Full Text] [PDF]

				W. M. Baarends, E. Wassenaar, J. W. Hoogerbrugge, G. van Cappellen, H. P. Roest, J. Vreeburg, M. Ooms, J. H. J. Hoeijmakers, and J. A. Grootegoed Loss of HR6B Ubiquitin-Conjugating Activity Results in Damaged Synaptonemal Complex Structure and Increased Crossing-Over Frequency during the Male Meiotic Prophase Mol. Cell. Biol., February 15, 2003; 23(4): 1151 - 1162. [Abstract] [Full Text] [PDF]

				L. S. Symington Role of RAD52 Epistasis Group Genes in Homologous Recombination and Double-Strand Break Repair Microbiol. Mol. Biol. Rev., December 1, 2002; 66(4): 630 - 670. [Abstract] [Full Text] [PDF]

				T. de los Santos, J. Loidl, B. Larkin, and N. M. Hollingsworth A Role for MMS4 in the Processing of Recombination Intermediates During Meiosis in Saccharomyces cerevisiae Genetics, December 1, 2001; 159(4): 1511 - 1525. [Abstract] [Full Text] [PDF]

				M. Lisby, R. Rothstein, and U. H. Mortensen From the Cover: Rad52 forms DNA repair and recombination centers during S phase PNAS, July 17, 2001; 98(15): 8276 - 8282. [Abstract] [Full Text] [PDF]

				A. J. R. Bishop, E. J. Louis, and R. H. Borts Minisatellite Variants Generated in Yeast Meiosis Involve DNA Removal During Gene Conversion Genetics, September 1, 2000; 156(1): 7 - 20. [Abstract] [Full Text]

				L. H. Rutkowski and R. E. Esposito Recombination Can Partially Substitute for SPO13 in Regulating Meiosis I in Budding Yeast Genetics, August 1, 2000; 155(4): 1607 - 1621. [Abstract] [Full Text]

				K. Suto, A. Nagata, H. Murakami, and H. Okayama A Double-Strand Break Repair Component Is Essential for S Phase Completion in Fission Yeast Cell Cycling Mol. Biol. Cell, October 1, 1999; 10(10): 3331 - 3343. [Abstract] [Full Text]

				F. K. Khasanov, G. V. Savchenko, E. V. Bashkirova, V. G. Korolev, W.-D. Heyer, and V. I. Bashkirov A New Recombinational DNA Repair Gene From Schizosaccharomyces pombe With Homology to Escherichia coli RecA Genetics, August 1, 1999; 152(4): 1557 - 1572. [Abstract] [Full Text]

				F. Paques and J. E. Haber Multiple Pathways of Recombination Induced by Double-Strand Breaks in Saccharomyces cerevisiae Microbiol. Mol. Biol. Rev., June 1, 1999; 63(2): 349 - 404. [Abstract] [Full Text] [PDF]

				S. L. Gasior, A. K. Wong, Y. Kora, A. Shinohara, and D. K. Bishop Rad52 associates with RPA and functions with Rad55 and Rad57 to assemble meiotic recombination complexes Genes & Dev., July 15, 1998; 12(14): 2208 - 2221. [Abstract] [Full Text]

				J. Engebrecht, S. Masse, L. Davis, K. Rose, and T. Kessel Yeast Meiotic Mutants Proficient for the Induction of Ectopic Recombination Genetics, February 1, 1998; 148(2): 581 - 598. [Abstract] [Full Text] [PDF]

				T. Wu and M Lichten Meiosis-induced double-strand break sites determined by yeast chromatin structure Science, January 28, 1994; 263(5146): 515 - 518. [Abstract] [PDF]

				G T Milne and D T Weaver Dominant negative alleles of RAD52 reveal a DNA repair/recombination complex including Rad51 and Rad52. Genes & Dev., September 1, 1993; 7(9): 1755 - 1765. [Abstract] [PDF]