Nuclease C Polymorphism of Calcium-Dependent Nucleases in Chlamydomonas reinhardtii |
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Authors: | Ogawa, Kazuo Kuroiwa, Tsuneyoshi |
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Affiliation: | National Institute for Basic Biology, Department of Cell Biology Okazaki 444, Japan |
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Abstract: | DNases were extracted from cells of Chlamydomonas reinhardtii.Their polymorphism and metal ion requirements were investigated.Most of the nucleases from this organism required Ca2+ for fullactivation; therefore, the name nuclease C has been given tothem. Zn2+ and Mn2+ restored activation, but their respectivepotencies were 1/8 and 1/32 that of Ca2+. An in situ nucleaseassay revealed that there are at least 6 species of nucleaseC. The molecular weights of the components were 26,000 (C1),25,000 (C2), 22,000 (C5), 21,000 (C6), 18,000 (C3) and 17,000(C4) by SDS PAGE. Ca2+-dependent nuclease activity was slightlyhigher in the female gamete than in the male gamete. There wasno marked change in the apparent nuclease activity during thefirst 3 h after mating. An in situ assay, however, showed thatnuclease C6 and C4 are present in smaller amounts in the malegamete in comparison to the female gamete, and that the contentsor activities of nuclease C5 and C6 diminish during the first3060 min after mating. This evidence is discussed interms of the possible participation of nuclease C in the maternalinheritance of chloroplast genes in this organism. (Received October 8, 1984; Accepted January 21, 1985) |
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