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转染的7SL RNA和Alu基因序列对HeLa细胞生长的调控

Modulation of HeLa cell growth by transfected 7SL RNA and Alu gene sequences.

作者信息

Sakamoto K, Fordis C M, Corsico C D, Howard T H, Howard B H

机构信息

Laboratory of Molecular Biology, National Cancer Institute, Bethesda, Maryland 20892.

出版信息

J Biol Chem. 1991 Feb 15;266(5):3031-8.

PMID:1704372
Abstract

Alu and 7SL RNA gene sequences were tested for the potential to regulate mammalian cell growth by introducing these sequences into HeLa cells in a coupled DEAE-dextran transfection/affinity cell sorting system. Both Alu and 7SL RNA genes mediated inhibition of [3H]thymidine and [35S]methionine incorporation in recipient cells. In addition, short term growth curves were performed on calcium phosphate/DNA cotransfected, affinity-purified cells. This second assay revealed that transfected Alu and 7SL RNA can also cause suppression of HeLa cell proliferation. To investigate whether transcription or polymerase III (pol III) transcription factor binding was required for inhibitory activity, mutations were introduced into RNA pol III B block promoter elements in each of these genes. Suppression of [3H]thymidine incorporation was dependent on the presence of this pol III element in both genes; likewise, 7SL RNA-mediated growth suppression required the presence of the pol III B block promoter element. The results of this study indicate that Alu and 7SL RNA gene sequences interact with cellular factors that are important for HeLa cell proliferation and suggest that these pol III-transcribed elements may be involved in the regulation of cellular growth.

摘要

通过在DEAE-葡聚糖转染/亲和细胞分选系统中将Alu和7SL RNA基因序列导入HeLa细胞,测试了它们调节哺乳动物细胞生长的潜力。Alu和7SL RNA基因均介导受体细胞中[³H]胸苷和[³⁵S]甲硫氨酸掺入的抑制。此外,对磷酸钙/DNA共转染、亲和纯化的细胞进行了短期生长曲线测定。这第二种测定方法表明,转染的Alu和7SL RNA也可导致HeLa细胞增殖的抑制。为了研究抑制活性是否需要转录或聚合酶III(pol III)转录因子结合,在这些基因的每个RNA pol III B阻断启动子元件中引入了突变。[³H]胸苷掺入的抑制取决于两个基因中该pol III元件的存在;同样,7SL RNA介导的生长抑制需要pol III B阻断启动子元件的存在。这项研究的结果表明,Alu和7SL RNA基因序列与对HeLa细胞增殖很重要的细胞因子相互作用,并表明这些pol III转录元件可能参与细胞生长的调节。

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