Interaction between Lim15/Dmc1 and the homologue of the large subunit of CAF-1: a molecular link between recombination and chromatin assembly during meiosis.
In eukaryotes, meiosis leads to genetically variable gametes through recombination between homologous chromosomes of maternal and paternal origin. Chromatin organization following meiotic recombination is critical to ensure the correct segregation of homologous chromosomes into gametes. However, the mechanism of chromatin organization after meiotic recombination is unknown. In this study we report that the meiosis-specific recombinase Lim15/Dmc1 interacts with the homologue of the largest subunit of chromatin assembly factor 1 (CAF-1) in the basidiomycete Coprinopsis cinerea (Coprinus cinereus). Using C. cinerea LIM15/DMC1 (CcLIM15) as the bait in a yeast two-hybrid screen, we have isolated the C. cinerea homologue of Cac1, the largest subunit of CAF-1 in Saccharomyces cerevisiae, and named it C. cinerea Cac1-like (CcCac1L). Two-hybrid assays confirmed that CcCac1L binds CcLim15 in vivo. beta-Galactosidase assays revealed that the N-terminus of CcCac1L preferentially interacts with CcLim15. Co-immunoprecipitation experiments showed that these proteins also interact in the crude extract of meiotic cells. Furthermore, we demonstrate that, during meiosis, CcCac1L interacts with proliferating cell nuclear antigen (PCNA), a component of the DNA synthesis machinery recently reported as an interacting partner of Lim15/Dmc1. Taken together, these results suggest a novel role of the CAF-1-PCNA complex in meiotic events. We propose that the CAF-1-PCNA complex modulates chromatin assembly following meiotic recombination.
Ishii
Satomi
S
Department of Applied Biological Science, Faculty of Science and Technology, Tokyo University of Science, Chiba, Japan.
Koshiyama
Akiyo
A
Hamada
Fumika N
FN
Nara
Takayuki Y
TY
Iwabata
Kazuki
K
Sakaguchi
Kengo
K
Namekawa
Satoshi H
SH
D018797:Cell Cycle Proteins / Q000378:metabolism*; D056504:Chromatin Assembly Factor-1; D042002:Chromatin Assembly and Disassembly; D002868:Chromosomal Proteins, Non-Histone / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D003302:Coprinus / Q000201:enzymology; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008540:Meiosis; D008954:Models, Biological; D018809:Proliferating Cell Nuclear Antigen / Q000235:genetics / Q000378:metabolism; D021122:Protein Subunits / Q000737:chemistry; D011995:Recombination, Genetic; D020349:Surface Plasmon Resonance; D020798:Two-Hybrid System Techniques
C533593:CHAF1B protein, human; D018797:Cell Cycle Proteins; D056504:Chromatin Assembly Factor-1; D002868:Chromosomal Proteins, Non-Histone; D004268:DNA-Binding Proteins; D018809:Proliferating Cell Nuclear Antigen; D021122:Protein Subunits; C074556:DMC1 protein, human