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Analysis of histone gene expression during the cell cycle in HeLa cells by using cloned human histone genes.

机译:使用克隆的人类组蛋白基因分析HeLa细胞在细胞周期中的组蛋白基因表达。

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摘要

Although it is generally agreed that histone protein synthesis is restricted to the S phase of the cell cycle--and therefore parallels DNA replication--both transcriptional and posttranscriptional levels of control have been invoked. Using blot hybridization with several cloned genomic human histone sequences representing different histone gene clusters as probes, we have assessed the steady-state level of histone RNAs in the nucleus and cytoplasm of G1 and S phase HeLa S3 cells. The representation of histone mRNA sequences of G1 compared with S phase cells was less than 1% in the cytoplasm and approximately 1% in the nucleus. These data are consistent with transcriptional control, but we cannot completely dismiss the possibility that regulation of histone gene expression is, to some extent, mediated posttranscriptionally. If histone gene transcription does occur in G1, the RNAs must either be rapidly degraded or be transcribed to a limited extent compared with S phase. An unexpected result was obtained when a blot of cytoplasmic RNA from G1 and S phase cells was hybridized with lambda HHG 41 DNA (containing H3 and H4 human genomic histone sequences). Although hybridization with histone mRNAs was observed for RNAs from S phase but not from G1 cells, hybridization with a nonhistone RNA of approximately 330 nucleotides present predominantly in G1 was also observed.
机译:尽管人们普遍同意,组蛋白的合成仅限于细胞周期的S期(因此平行于DNA复制),但已调用了转录和转录后水平的控制。使用印迹杂交与代表不同组蛋白基因簇的几个克隆的基因组人组蛋白序列作为探针,我们评估了G1和S期HeLa S3细胞核和细胞质中组蛋白RNA的稳态水平。与S期细胞相比,G1组蛋白mRNA序列的表达在细胞质中少于1%,在细胞核中少于1%。这些数据与转录控制一致,但是我们不能完全排除组蛋白基因表达的调节在某种程度上是转录后介导的可能性。如果G1中确实发生了组蛋白基因转录,则与S期相比,RNA必须迅速降解或转录到一定程度。当来自G1期和S期细胞的细胞质RNA印迹与λHHG 41 DNA(含有H3和H4人基因组蛋白序列)杂交时,获得了出乎意料的结果。尽管对于来自S期而不是来自G1细胞的RNA观察到与组蛋白mRNA的杂交,但也观察到与G1细胞中主要存在的约330个核苷酸的非组蛋白RNA杂交。

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