Nucleic Acids Research, 2000, Vol. 28, No. 4 932-939
© 2000 Oxford University Press
Structure and expression of the human p68 RNA helicase gene
Medizinische Biochemie und Molekularbiologie, Universität des Saarlandes, 66421 Homburg, Germany
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.
* To whom correspondence should be addressed. Tel: +49 6841 16 6020; Fax: +49 6841 16 6521; Email: bchsta@med-rz.uni-sb.de Present addresses: Peter Hloch, Roche Molecular Biochemicals, Nonnenwald 2, D-82372 Penzberg, Germany Thomas Weitzenegger, Department of Molecular Biology, Byk Gulden Pharmaceuticals, D-78434 Konstanz, Germany
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