Nucleic Acids Research, Vol 24, Issue 23 4684-4692, Copyright © 1996 by Oxford University Press
SA Matis, K Martincic and C Milcarek
Early/memory and plasma B-cell lines and fibroblasts were analyzed for
their ability to use a 5' proximal (variant) versus a 3' distal (constant)
poly(A) site, in the absence of a competing splice, from a set of related
constructs. The proximal:distal poly(A) site use (P:D ratio) of the
resulting cytoplasmic poly(A)+ mRNA is a measure of poly(A) site strength.
In this context the immunoglobulin gamma2b secretory-specific poly(A) site
showed a P:D ratio of 1:1 in plasma cells, 0.43:1 in early/memory B-cells
and an intermediate value in fibroblasts. Meanwhile, a construct with a
proximal SV40 early-like poly(A) site produced mRNA with a P:D ratio of
>>50:1 in all cell types. Alterations in the region downstream of the
proximal poly(A) addition site and at the site itself resulted in changes
in the P:D ratio. However, these poly(A) sites, all with a P:D ratio of
< or = 5:1, were used most efficiently in plasma cells. Constructs
totally devoid of immunoglobulin sequences, but containing heterologous
poly(A) sites producing mRNA with P:D ratios of < or = 5:1, were also
used more efficiently in plasma cells. We therefore conclude that weak
poly(A) sites, regardless of sequence composition, are used more
efficiently in plasma cells than in the other cell types.
ARTICLES
B-lineage regulated polyadenylation occurs on weak poly(A) sites regardless of sequence composition at the cleavage and downstream regions
Department of Molecular Genetics and Biochemistry, University of Pittsburgh School of Medicine, PA 15261-2072, USA.
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