Transcription and polyadenylation in a short human intergenic region.

The poly(A) signal of the human Lamin B2 gene was previously shown to lie 600 bp upstream of the cap site of a gene of unknown function (ppv 1). However, using RNase protection analysis, we show that ppv 1 has two clusters of multiple initiation sites, so that the 5"cap site lies only approxima...

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Main Authors: Brackenridge, S, Ashe, H, Giacca, M, Proudfoot, N
Format: Journal article
Language:English
Published: 1997
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author Brackenridge, S
Ashe, H
Giacca, M
Proudfoot, N
author_facet Brackenridge, S
Ashe, H
Giacca, M
Proudfoot, N
author_sort Brackenridge, S
collection OXFORD
description The poly(A) signal of the human Lamin B2 gene was previously shown to lie 600 bp upstream of the cap site of a gene of unknown function (ppv 1). However, using RNase protection analysis, we show that ppv 1 has two clusters of multiple initiation sites, so that the 5"cap site lies only approximately 280 nt downstream of the Lamin B2 poly(A) signal. We analysed nascent transcription across this unusually short intergenic region using nuclear run-on analysis of both the endogenous locus and of transiently transfected hybrid constructs. Surprisingly, transcription of the Lamin B2 gene does not appear to terminate prior to any of the mapped ppv 1 start sites, although pausing of the elongating polymerase complexes is observed downstream of the Lamin B2 poly(A) signal. We suggest that this pausing may be sufficient to protect the downstream gene from transcriptional interference. Finally, we have also investigated the sequences required for efficient recognition of the Lamin B2 poly(A) signal. We show that sequences upstream of the AAUAAA element are required for full activity, which is an unusual feature of mammalian poly(A) signals.
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spelling oxford-uuid:834b54b1-7293-42dd-b396-0ee59bbbc9da2022-03-26T21:43:14ZTranscription and polyadenylation in a short human intergenic region.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:834b54b1-7293-42dd-b396-0ee59bbbc9daEnglishSymplectic Elements at Oxford1997Brackenridge, SAshe, HGiacca, MProudfoot, NThe poly(A) signal of the human Lamin B2 gene was previously shown to lie 600 bp upstream of the cap site of a gene of unknown function (ppv 1). However, using RNase protection analysis, we show that ppv 1 has two clusters of multiple initiation sites, so that the 5"cap site lies only approximately 280 nt downstream of the Lamin B2 poly(A) signal. We analysed nascent transcription across this unusually short intergenic region using nuclear run-on analysis of both the endogenous locus and of transiently transfected hybrid constructs. Surprisingly, transcription of the Lamin B2 gene does not appear to terminate prior to any of the mapped ppv 1 start sites, although pausing of the elongating polymerase complexes is observed downstream of the Lamin B2 poly(A) signal. We suggest that this pausing may be sufficient to protect the downstream gene from transcriptional interference. Finally, we have also investigated the sequences required for efficient recognition of the Lamin B2 poly(A) signal. We show that sequences upstream of the AAUAAA element are required for full activity, which is an unusual feature of mammalian poly(A) signals.
spellingShingle Brackenridge, S
Ashe, H
Giacca, M
Proudfoot, N
Transcription and polyadenylation in a short human intergenic region.
title Transcription and polyadenylation in a short human intergenic region.
title_full Transcription and polyadenylation in a short human intergenic region.
title_fullStr Transcription and polyadenylation in a short human intergenic region.
title_full_unstemmed Transcription and polyadenylation in a short human intergenic region.
title_short Transcription and polyadenylation in a short human intergenic region.
title_sort transcription and polyadenylation in a short human intergenic region
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AT asheh transcriptionandpolyadenylationinashorthumanintergenicregion
AT giaccam transcriptionandpolyadenylationinashorthumanintergenicregion
AT proudfootn transcriptionandpolyadenylationinashorthumanintergenicregion