Hop2-Mnd1 condenses DNA to stimulate the synapsis phase of DNA strand exchange.
Hop2-Mnd1 is a meiotic recombination mediator that stimulates DNA strand invasion by both Dmc1 and Rad51. To understand the biochemical mechanism of this stimulation, we directly visualized the heterodimer acting on single molecules of duplex DNA using optical tweezers and video fluorescence microscopy. The results show that the Hop2-Mnd1 heterodimer efficiently condenses double-stranded DNA via formation of a bright spot or DNA condensate. The condensation of DNA is Hop2-Mnd1 concentration-dependent, reversible, and specific to the heterodimer, as neither Hop2 nor Mnd1 acting alone can facilitate this reaction. The results also show that the rate-limiting nucleation step of DNA condensation is overcome in the presence of divalent metal ions, with the following order of preference: Mn(2+)>Mg(2+)>Ca(2+). Hop2-Mnd1/Dmc1/single-stranded DNA nucleoprotein filaments also condense double-stranded DNA in a heterodimer concentration-dependent manner. Of importance, the concentration dependence parallels that seen in DNA strand exchange. We propose that rapid DNA condensation is a key factor in stimulating synapsis, whereas decondensation may facilitate the invasion step and/or the ensuing branch migration process.
Genetics and Biochemistry Branch, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland, USA.
Roberto J
RJ
Pezza
R Daniel
RD
Camerini-Otero
Piero R
PR
Bianco
D000818:Animals; D002413:Cations, Divalent / Q000494:pharmacology; D018797:Cell Cycle Proteins / Q000378:metabolism*; D023902:Chromosome Pairing / Q000187:drug effects; D004247:DNA / Q000737:chemistry* / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism; D006801:Humans; D051379:Mice; D008958:Models, Molecular; D009690:Nucleic Acid Conformation
D002413:Cations, Divalent; D018797:Cell Cycle Proteins; D004268:DNA-Binding Proteins; C501203:Mnd1 protein, mouse; D004247:DNA; C074556:DMC1 protein, human