Sakiyama S, Yoda K
Gan To Kagaku Ryoho. 1983 Feb;10(2 Pt 2):539-43.
Cytoplasmic isoactins of mouse fibroblast L-cells were examined by two-dimensional gel electrophoresis. In contrast to other cultured cell lines, which contain both beta- and gamma-actins. L-cells contained only beta-actin species. When RNA of L-cells were translated in a nuclease-treated reticulocyte lysate system, actin synthesized in vitro also appeared as only beta-actin. After digestion of genomic DNAs of both L-cells and mouse brain, which contains beta- and gamma-actins, by Eco RI, Hind III, Bam HI and Sal I, obtained fragments were investigated by Southern blott analysis using 32P -labeled probe constructed from PA1 which carries a full length copy of chicken beta-actin mRNA. No differences in the patterns were detected between L-cell and brain DNAs. This result suggests that the absence of gamma-actin expression in L-cells is not due to gross deletions, insertions or rearrangements of isoactin genes. However, the use of the isoschizomeric pair of restriction enzymes, Hpa II and Msp I, resulted in different patterns of isoactin genes, implying a possible involvement of methylation in regulation of isoactin gene expression.
通过二维凝胶电泳对小鼠成纤维细胞L-细胞的细胞质同工肌动蛋白进行了检测。与其他同时含有β-肌动蛋白和γ-肌动蛋白的培养细胞系不同,L-细胞仅含有β-肌动蛋白。当L-细胞的RNA在经核酸酶处理的网织红细胞裂解物系统中进行翻译时,体外合成的肌动蛋白也仅表现为β-肌动蛋白。在用Eco RI、Hind III、Bam HI和Sal I对同时含有β-肌动蛋白和γ-肌动蛋白的L-细胞和小鼠脑的基因组DNA进行消化后,使用由携带鸡β-肌动蛋白mRNA全长拷贝的PA1构建的32P标记探针,通过Southern印迹分析对获得的片段进行了研究。未检测到L-细胞和脑DNA之间的模式差异。该结果表明,L-细胞中γ-肌动蛋白表达的缺失并非由于同工肌动蛋白基因的大片段缺失、插入或重排。然而,使用同裂酶对Hpa II和Msp I,导致了同工肌动蛋白基因的不同模式,这意味着甲基化可能参与了同工肌动蛋白基因表达的调控。