Cookies on this website
We use cookies to ensure that we give you the best experience on our website. If you click 'Continue' we'll assume that you are happy to receive all cookies and you won't see this message again. Click 'Find out more' for information on how to change your cookie settings.

The multicopy human U2 small nuclear (sn)RNA genes are transcribed by RNA polymerase (pol) II and contain two major promoter elements upstream of the transcription start site: an essential proximal sequence element (PSE) at around -55 and a distal sequence element (DSE) at around -220. We have carried out an in vivo footprinting analysis on these genes, and the results suggest that most, if not all, of the U2 gene promoters are bound by factors in interphase. Both the DSE and the PSE are protected from digestion, and the pattern of methylation protection over the DSE is virtually identical to that obtained in vitro using nuclear extract. Our results also indicate that the DNA between the PSE and the transcription start site is distorted and that proteins interact with the promoter between -20 and -33. Mutation of this sequence affects both the accuracy of initiation and polymerase specificity, underlining the importance of this region in U2 gene expression. We have also analysed the pattern of protection over the DSE and PSE of the U2 genes in mitotic cells. The degree of protection over all promoter elements is drastically reduced, suggesting that loss of DNA binding factors from the promoter plays a role in the shutdown of U2 gene transcription in mitosis.

Type

Journal article

Journal

Gene

Publication Date

02/10/2003

Volume

315

Pages

103 - 112

Keywords

Base Sequence, Binding Sites, DNA Footprinting, DNA Methylation, DNA-Directed RNA Polymerases, HeLa Cells, Humans, Mitosis, Models, Genetic, Mutation, Promoter Regions, Genetic, Protein Binding, RNA, Small Nuclear, Regulatory Sequences, Nucleic Acid, Substrate Specificity, Transcription Factors, Transcription Initiation Site, Transcription, Genetic