Nucleic Acids Research, Vol 25, Issue 12 2326-2336, Copyright © 1997 by Oxford University Press
S Brackenridge, HL Ashe, M Giacca and NJ Proudfoot
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.
ARTICLES
Transcription and polyadenylation in a short human intergenic region
Sir William Dunn School of Pathology, University of Oxford, South Parks Road, Oxford OX1 3RE, UK.
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