Kölling R, Nguyen T, Chen E Y, Botstein D
Department of Cell Genetics, Genentech Inc., South San Francisco, CA 94305.
Mol Gen Genet. 1993 Mar;237(3):359-69. doi: 10.1007/BF00279439.
We isolated a gene encoding a 218 kDa myosin-like protein from Saccharomyces cerevisiae using a monoclonal antibody directed against human platelet myosin as a probe. The protein sequence encoded by the MLP1 gene (for myosin-like protein) contains extensive stretches of a heptad-repeat pattern suggesting that the protein can form coiled coils typical of myosins. Immunolocalization experiments using affinity-purified antibodies raised against a TrpE-MLP1 fusion protein showed a dot-like structure adjacent to the nucleus in yeast cells bearing the MLP1 gene on a multicopy plasmid. In mouse epithelial cells the yeast anti-MLP1 antibodies stained the nucleus. Mutants bearing disruptions of the MLP1 gene were viable, but more sensitive to ultraviolet light than wild-type strains, suggesting an involvement of MLP1 in DNA repair. The MLP1 gene was mapped to chromosome 11, 25 cM from met1.
我们使用针对人血小板肌球蛋白的单克隆抗体作为探针,从酿酒酵母中分离出一个编码218 kDa肌球蛋白样蛋白的基因。MLP1基因(肌球蛋白样蛋白基因)编码的蛋白质序列包含大量七肽重复模式的片段,这表明该蛋白可以形成典型的肌球蛋白卷曲螺旋结构。使用针对TrpE-MLP1融合蛋白产生的亲和纯化抗体进行的免疫定位实验表明,在多拷贝质粒上携带MLP1基因的酵母细胞中,有一个点状结构毗邻细胞核。在小鼠上皮细胞中,酵母抗MLP1抗体对细胞核进行了染色。携带MLP1基因破坏突变的菌株是可存活的,但比野生型菌株对紫外线更敏感,这表明MLP1参与DNA修复。MLP1基因被定位到11号染色体上,距离met1基因25 cM。