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Premature termination of RNA polymerase II mediated transcription of a seed protein gene in Schizosaccharomyces pombe

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Title Premature termination of RNA polymerase II mediated transcription of a seed protein gene in Schizosaccharomyces pombe
 
Creator Chakraborty, Subhra
Sarmah, Bhaskarjyoti
Chakraborty, Niranjan
Datta, Asis
 
Subject Schizosaccharomyces pombe
RNA polymerase II mediated transcription
seed protein gene
 
Description The poly(A) signal and downstream elements with
transcriptional pausing activity play an important
role in termination of RNA polymerase II transcription.
We show that an intronic sequence derived from the
plant seed protein gene (AmA1 ) specifically acts as a
transcriptional terminator in the fission yeast, Schizosaccharomyces pombe. The 3′-end points of mRNA
encoded by the AmA1 gene were mapped at different
positions in S.pombe and in native cells of Amaranthus
hypochondriacus. Deletion analyses of the AmA1 intronic sequence revealed that multiple elements
essential for proper transcriptional termination in
S.pombe include two site-determining elements
(SDEs) and three downstream sequence elements.
RT–PCR analyses detected transcripts up to the
second SDE. This is the first report showing that the
highly conserved mammalian poly(A) signal,
AAUAAA, is also functional in S.pombe. The poly(A)
site was determined as Y(A) both in native and heterologous systems but at different positions. Deletion of
these cis-elements abolished 3′-end processing in
S.pombe and a single point mutation in this motif reduced the activity by 70% while enhancing activity
at downstream SDE. These results indicate that the
bipartite sequence elements as signals for 3′-end
processing in fission yeast act in tandem with other
cis-acting elements. A comparison of these elements
in the AmA1 intron that function as a transcriptional
terminator in fission yeast with that of its native
genes showed that both require an AT-rich distal and
proximal upstream element. However, these
sequences are not identical. Transcription run-on
analysis indicates that elongating RNA polymerase II
molecules accumulate over these pause signals,
maximal at 611–949 nt. Furthermore, we demonstrate
that the AmA1 intronic terminator sequence acts in a position-independent manner when placed within
another gene.
This work was supported by a grant from the
Department of Biotechnology, Ministry of Science and
Technology, Government of India.
 
Date 2013-10-18T07:07:09Z
2013-10-18T07:07:09Z
2002
2 April 2002
 
Type Article
 
Identifier Nucl. Acids Res., 30: 2940-2949
http://hdl.handle.net/123456789/25
 
Language en
 
Publisher Oxford University Press