Strand exchange reaction in vitro and DNA-dependent ATPase activity of recombinant LIM15/DMC1 and RAD51 proteins from Coprinus cinereus.
We previously cloned recA-homolog genes from a basidiomycete, Coprinus cinereus, and obtained the recombinant proteins (Nara et al., Mol. Gen. Genet. 262, 781-789, 1999, see Ref. 1; Nara and Sakaguchi, Biochem. Biophys. Res. Commun. 275, 97-102, 2000, see Ref. 2). The primary purpose of the present study was to characterize the biochemical properties of the recombinant LIM15/DMC1 (CoLIM15) and RAD51 (CoRAD51) proteins. We purified the recombinant proteins, and their molecular masses were 37 and 35 kDa, respectively. Both enzymes showed DNA-dependent ATPase activity and ATP-dependent strand exchange reaction in vitro. CoRad51 was a five- to sixfold stronger DNA-dependent ATPase and showed greater dependency on single-stranded DNA than CoLim15. In meiosis, both enzymes were highly accumulated in the meiotic tissue at leptotene and zygotene stages at which the homologous chromosomes pair, but disappeared just before the pachytene stage at which they recombine. From these and the previously reported results, we discuss here the relationships between the enzymes and meiosis.
Nara
T
T
Department of Applied Biological Science, Faculty of Science and Technology, Science University of Tokyo, 2641 Yamazaki, Chiba-ken, Noda-shi, 278, Japan.
Hamada
F
F
Namekawa
S
S
Sakaguchi
K
K
D000251:Adenosine Triphosphatases / Q000378:metabolism*; D001487:Basidiomycota / Q000201:enzymology*; D015153:Blotting, Western; D018797:Cell Cycle Proteins; D004265:DNA Helicases / Q000378:metabolism*; D004277:DNA, Single-Stranded / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism*; D005656:Fungal Proteins / Q000378:metabolism*; D066298:In Vitro Techniques; D051135:Rad51 Recombinase; D011994:Recombinant Proteins / Q000378:metabolism
D018797:Cell Cycle Proteins; D004277:DNA, Single-Stranded; D004268:DNA-Binding Proteins; D005656:Fungal Proteins; D011994:Recombinant Proteins; D051135:Rad51 Recombinase; D000251:Adenosine Triphosphatases; D004265:DNA Helicases