Rössler O G, Hloch P, Schütz N, Weitzenegger T, Stahl H
Medizinische Biochemie und Molekularbiologie, Universität des Saarlandes, 66421 Homburg, Germany.
Nucleic Acids Res. 2000 Feb 15;28(4):932-9. doi: 10.1093/nar/28.4.932.
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.
核DEAD盒蛋白p68与SV40大肿瘤抗原在免疫上相关,且这两种蛋白都具有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相关方面。Northern印迹和引物延伸分析表明,尽管其水平存在差异,但p68 RNA解旋酶似乎在所有测试组织中均从单一转录起始位点表达。序列分析显示,p68启动子含有一个“TATA”盒、一个“CAAT”盒和一个起始子元件,并包含Sp1、AP - 2、CRE和Myc的高亲和力结合位点。这以及瞬时表达试验中的功能性启动子分析表明,p68基因的转录调控很复杂。此外,有迹象表明p68的表达在转录后也受到调控。来自人类细胞的聚腺苷酸(poly(A))⁺RNA的稳态库包含完全剪接的p68 mRNA以及包含第8 - 11或8 - 12号内含子(总共12个内含子)的可变剪接形式,这些形式不会被翻译。对一个条件性过表达p68的HeLa细胞系的分析表明在剪接水平存在负向自我调节,这一点已被最近报道的p68与人细胞中的剪接体的关联所证实。