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Distinct roles of two separable in vitro activities of yeast Mre11 in mitotic and meiotic recombination
Author(s) -
Furuse Munenori,
Nagase Yuko,
Tsubouchi Hideo,
MurakamiMurofushi Kimiko,
Shibata Takehiko,
Ohta Kunihiro
Publication year - 1998
Publication title -
the embo journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 7.484
H-Index - 392
eISSN - 1460-2075
pISSN - 0261-4189
DOI - 10.1093/emboj/17.21.6412
Subject(s) - biology , meiosis , mitotic crossover , yeast , mitosis , recombination , genetics , microbiology and biotechnology , genetic recombination , homologous recombination , in vitro , ploidy , saccharomyces cerevisiae , gene
In Saccharomyces cerevisiae , Mre11 protein is involved in both double‐strand DNA break (DSB) repair and meiotic DSB formation. Here, we report the correlation of nuclease and DNA‐binding activities of Mre11 with its functions in DNA repair and meiotic DSB formation. Purified Mre11 bound to DNA efficiently and was shown to have Mn 2+ ‐dependent nuclease activities. A point mutation in the N‐terminal phosphoesterase motif (Mre11D16A) resulted in the abolition of nuclease activities but had no significant effect on DNA binding. The wild‐type level of nuclease activity was detected in a C‐terminal truncated protein (Mre11ΔC49), although it had reduced DNA‐binding activity. Phenotypes of the corresponding mutations were also analyzed. The mre11D16A mutation conferred methyl methanesulfonate‐sensitivity to mitotic cells and caused the accumulation of unprocessed meiotic DSBs. The mre11ΔC49 mutant exhibited almost wild‐type phenotypes in mitosis. However, in meiosis, no DSB formation could be detected and an aberrant chromatin configuration was observed at DSB sites in the mre11ΔC49 mutant. These results indicate that Mre11 has two separable functional domains: the N‐terminal nuclease domain required for DSB repair, and the C‐terminal dsDNA‐binding domain essential to its meiotic functions such as chromatin modification and DSB formation.