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Cell. 1990 Jul 13;62(1):25-34.

A protein factor, ASF, controls cell-specific alternative splicing of SV40 early pre-mRNA in vitro.

Author information

1
Department of Biological Sciences, Columbia University, New York, New York 10027.

Abstract

SV40 early pre-mRNA is alternatively spliced by utilization of two different 5' splice sites and a shared 3' splice site to produce large T and small t mRNAs. The ratio of small t to large T mRNAs produced in human embryonic kidney 293 cells is 10- to 20-fold greater than in other mammalian cells, suggesting the existence of a 293 cell-specific factor that modulates alternative splicing. Here we show that nuclear extracts from 293 cells give rise to significantly more small t splicing than do extracts from HeLa cells. Using an in vitro complementation assay, we have characterized and extensively purified a factor from 293 extracts that brings about striking increases in small t splicing with concomitant decreases in large T splicing. The factor is heat sensitive and micrococcal nuclease resistant, suggesting that it is a protein lacking an accessible RNA component. Purification of the alternative splicing factor indicates that the activity is contained in one of several possibly related polypeptides of 30-35 kd.

PMID:
2163768
DOI:
10.1016/0092-8674(90)90236-8
[Indexed for MEDLINE]

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