Actin mRNAs in HeLa cells. Stabilization after adenovirus infection.
Actin mRNAs in HeLa cells. Stabilization after adenovirus infection.
复制标题
HeLa 细胞中的肌动蛋白 mRNA。
DOI：
10.1016/0022-2836(84)90268-7
复制
发表时间：
1984
影响因子：
5.6
通讯作者：
Weinmann,R
中科院分区：
文献类型：
--
作者：
Khalili,K;Weinmann,R
我们对HeLa细胞和腺病毒感染的HeLa细胞中肌动蛋白基因的表达进行了比较分析。在这些细胞中，通过对新合成的RNA进行脉冲标记和/或在从未感染或感染的HeLa细胞分离的细胞核中进行体外转录来检测肌动蛋白基因转录率。此外，通过斑点和Northern blot检测了肌动蛋白特异性异质核RNA的积累和肌动蛋白mrna在细胞质中的出现率。2型腺病毒感染HeLa细胞后，肌动蛋白基因转录率保持不变，细胞核内肌动蛋白前体水平略有降低。在感染细胞中，新合成的肌动蛋白mRNA以非常低的速率进入细胞质。转运不足不影响细胞质中信息的稳态水平。通过传统的脉冲追踪实验和使用不依赖于标记RNA的5-6-diCl-1-β-d-核糖呋喃基苯并咪唑的新方法分析细胞质肌动蛋白mrna的半衰期。两种方法的结果相同。未感染的HeLa细胞具有半衰期相对较短的肌动蛋白mrna，从不到6到12小时不等。相比之下，在腺病毒感染细胞的细胞质中，肌动蛋白特异性mrna的半衰期大于14至24小时。这些观察结果表明，尽管在腺病毒感染后肌动蛋白mrna向细胞质的转运速率降低，但增加的半衰期导致肌动蛋白mrna在细胞质中积累到正常水平。
We have carried out a comparative analysis of the expression of the actin genes in HeLa and adenovirus-infected HeLa cells. The rate of actin gene transcription was examined in these cells by pulse-labeling of the newly synthesized RNA and/or byin vitrotranscription in nuclei isolated from uninfected or infected HeLa cells. In addition, accumulation of actin-specific heterogeneous nuclear RNA, and rate of appearance of the actin mRNAs in the cytoplasm were examined by dot and Northern blot analysis. The rate of actin gene transcription remained constant after infection of HeLa cells with adenovirus serotype 2, while the level of the actin precursor in the nuclei was slightly reduced. In the infected cells, newly synthesized actin mRNA enters the cytoplasm at a very reduced rate. The deficiency of transport does not affect the steady-state level of the messages in the cytoplasm. The half-life of cytoplasmic actin mRNAs was analyzed by traditional pulse-chase experiments and by a novel procedure using 5-6-diCl-1-β-d-ribofuranosylbenzimidazole, which does not rely on labeled RNA. Both procedures gave identical results. Uninfected HeLa cells have actin mRNAs with relatively short half-lives, from less than six to 12 hours. In contrast, the half-lives of the actin-specific mRNAs, in the cytoplasm of adenovirus-infected cells, is greater than 14 to 24 hours. These observations suggest that, although the rate of transport of actin mRNAs to the cytoplasm is reduced upon infection with adenovirus, increased half-lives result in accumulation of actin mRNAs to normal levels in the cytoplasm.