Ryseck R P, MacDonald-Bravo H, Zerial M, Bravo R
European Molecular Biology Laboratory, Heidelberg, Federal Republic of Germany.
Exp Cell Res. 1989 Feb;180(2):537-45. doi: 10.1016/0014-4827(89)90080-3.
From a collection of more than 80 nonoverlapping clones, isolated by differential screening of a lambda cDNA library prepared from serum-stimulated cells in the presence of cycloheximide, we have identified four clones that encoded for components of the cytoskeleton and extracellular matrix. DNA sequencing of clones B2, V58, TT1, and P38 demonstrated that they corresponded to beta-actin, alpha-tropomyosin, fibronectin, and the beta-subunit of fibronectin receptor. All four mRNA levels showed a detectable increase 30 min after stimulation and remained at high levels for at least 8 h. The half-lives of these mRNAs were found to be very long in contrast to those of other growth factor-inducible genes. An increase in transcription was observed for the four genes. Actin and fibronectin showed nearly maximal increase at 15 min, while fibronectin receptor and tropomyosin reached their maximum transcription at 1 h. These results demonstrated that four interacting components of the cytoskeleton and extracellular matrix are rapidly induced in stimulated quiescent cells, possibly reflecting part of the coordinate changes in gene expression that occur during embryogenesis and wound healing.
从80多个非重叠克隆的集合中,通过对在放线菌酮存在下由血清刺激细胞制备的λ cDNA文库进行差异筛选分离得到,我们鉴定出四个编码细胞骨架和细胞外基质成分的克隆。对克隆B2、V58、TT1和P38进行DNA测序表明,它们分别对应于β - 肌动蛋白、α - 原肌球蛋白、纤连蛋白和纤连蛋白受体的β亚基。所有四种mRNA水平在刺激后30分钟均显示出可检测到的增加,并在至少8小时内保持高水平。与其他生长因子诱导基因的mRNA半衰期相比,发现这些mRNA的半衰期非常长。观察到这四个基因的转录增加。肌动蛋白和纤连蛋白在15分钟时显示出几乎最大的增加,而纤连蛋白受体和原肌球蛋白在1小时时达到最大转录。这些结果表明,细胞骨架和细胞外基质的四个相互作用成分在受刺激的静止细胞中被快速诱导,这可能反映了胚胎发育和伤口愈合过程中发生的基因表达协调变化的一部分。