Perriard J C
J Biol Chem. 1979 Aug 10;254(15):7036-41.
During the first 2 days of culture of myogenic chicken cells only minute amounts of mRNA for a subunit of creatine kinase, M-CK, could be found in cultures from the 3rd and 4th day; however, a dramatic increase of mRNA for the muscle-specific M-CK was observed. The mRNA for B-CK, another subunit of creatine kinase, was readily detectable in young cultures increased up to the 3rd day in culture but decreased significantly 1 day later. The comparison of the ratios of mRNA concentrations for M-CK and B-CK in polysomal and total cellular RNA did not reveal significant differences in polysomal and total cellular RNA from 24-, 48-, and 68-h cultures, indicating that specific accumulation of a nonpolysomal form containing translatable mRNA for M-CK prior to its appearance in the polysomes does not seem to represent a major mechanism of regulation. In RNA from cells grown in 5-bromo-2'-deoxyuridine (BrdUrd) for 3 days and subcultured into the same medium for another 3 days only mRNA for B-CK was found. If cells, however, were subcultured into standard medium both mRNAs, the one for B-CK and the one for M-CK could be detected. BrdUrd inhibition seems to affect only the mRNA for M-CK and not the one for B-CK. Removal of BrdUrd from the cultures probably allows new transcription or processing of mRNA for M-CK, or both.
在生肌鸡细胞培养的头两天,仅能在培养物中检测到微量的肌酸激酶一个亚基(M-CK)的mRNA;从第三天和第四天起,可观察到肌肉特异性M-CK的mRNA急剧增加。肌酸激酶的另一个亚基B-CK的mRNA在年轻培养物中易于检测到,在培养至第三天时增加,但在一天后显著下降。对多核糖体RNA和总细胞RNA中M-CK与B-CK的mRNA浓度比值进行比较,未发现24小时、48小时和68小时培养物的多核糖体RNA和总细胞RNA存在显著差异,这表明在M-CK的可翻译mRNA出现在多核糖体之前,含有该mRNA的非多核糖体形式的特异性积累似乎并非主要的调控机制。在含有5-溴-2'-脱氧尿苷(BrdUrd)的培养基中生长3天并再转接至相同培养基中培养3天的细胞的RNA中,仅发现了B-CK的mRNA。然而,如果将细胞转接至标准培养基中,则可检测到B-CK和M-CK的mRNA。BrdUrd抑制似乎仅影响M-CK的mRNA,而不影响B-CK的mRNA。从培养物中去除BrdUrd可能会允许M-CK的mRNA进行新的转录或加工,或者两者皆有。