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Structure and expression of the human p68 RNA helicase gene.

机译:人p68 RNA解旋酶基因的结构和表达。

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Nuclear DEAD box protein p68 is immunologically related to SV40 large tumour antigen and both proteins possess RNA helicase activity. In this report, we describe the structural organisation of the human p68 gene and aspects of the regulation of its expression. Northern blot and primer extension analyses indicate that, although its level is variable, the p68 RNA helicase appears to be expressed from a single transcription start site in all tissues tested. Sequence analysis revealed that the p68 promoter harbours a 'TATA', a 'CAAT' and an initiator element and contains high affinity binding sites for Sp1, AP-2, CRE and Myc. This and functional promoter analyses in transient expression assays suggest that transcriptional regulation of the p68 gene is complex. Furthermore, there are indications that p68 expression is also regulated post-transcriptionally. Steady-state pools of poly(A)(+)RNA from human cells contain completely spliced p68 mRNA and alternatively spliced forms that contain introns 8-11 or 8-12 (from a total of 12 introns) and are not translated. Analysis of a conditionally p68-overproducing HeLa cell line points to negative autoregulation at the level of splicing, which is confirmed by a recently reported association of p68 with spliceosomes in human cells.
机译:核DEAD盒蛋白p68与SV40大肿瘤抗原在免疫学上相关,并且两种蛋白均具有RNA解旋酶活性。在本报告中,我们描述了人类p68基因的结构组织及其表达调控的方面。 Northern印迹和引物延伸分析表明,尽管其水平可变,但p68 RNA解旋酶似乎在所有测试的组织中均从单个转录起始位点表达。序列分析表明,p68启动子带有一个“ TATA”,一个“ CAAT”和一个启动子元件,并含有对Sp1,AP-2,CRE和Myc的高亲和力结合位点。瞬时表达测定中的这种和功能性启动子分析表明,p68基因的转录调控是复杂的。此外,有迹象表明,转录后p68的表达也受到调节​​。来自人细胞的poly(A)(+)RNA的稳态库包含完全剪接的p68 mRNA和其他剪接形式,其中包含内含子8-11或8-12(来自总共12个内含子)并且不翻译。有条件地生产p68的HeLa细胞系的分析表明,在剪接水平存在负的自调控作用,这一点已被最近报道的人细胞中p68与剪接体的关联所证实。

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