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J. Biol. Chem., Vol. 276, Issue 33, 31004-31015, August 17, 2001
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From the The mammalian 70K protein, a component of
the U1 small nuclear ribonucleoprotein involved in pre-mRNA
splicing, interacts with a number of proteins important for regulating
constitutive and alternative splicing. Similar proteins that interact
with the yeast homolog of the 70K protein, Snp1p, have yet to be
identified. We used the two-hybrid system to find four
U1-Snp1 associating (Usa) proteins.
Two of these proteins physically associate with Snp1p as assayed by
coimmunoprecipitation. One is Prp8p, a known, essential spliceosomal
component. This interaction suggests some novel functions for Snp1p and
the U1 small nuclear ribonucleoprotein late in spliceosome development.
The other, Exo84p, is a conserved subunit of the exocyst, an
eight-protein complex functioning in secretion. We show here that
Exo84p is also involved in pre-mRNA splicing. A
temperature-sensitive exo84 mutation caused increased ratios of pre-mRNA to mRNA for the Rpl30 and actin transcripts in cells incubated at the non-permissive temperature. The mutation also
led to a defect in splicing and prespliceosome formation in
vitro; an indication that Exo84p has a direct role in splicing. The results elucidate a surprising link between splicing and secretion.
New Roles for the Snp1 and Exo84 Proteins in Yeast Pre-mRNA
Splicing*
§,¶,,
, and**
Department of Molecular Genetics and
Microbiology, University of New Mexico Health Sciences Center,
Cancer Research and Treatment Center,
Albuquerque, New Mexico 87131
*
This work was supported by National Science Foundation
Grants MCB9219408 and MCB9709915 and by grants from the Dedicated
Health Research Funds Committee of the University of New Mexico Health Sciences Center.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
Current address: Lovelace Respiratory Research Institute,
Albuquerque, NM 87185-5890.
**
To whom correspondence should be addressed: Dept. of Molecular
Genetics and Microbiology, University of New Mexico Health Sciences
Center, 900 Camino de Salud, NE, Albuquerque, NM 87131. Tel.:
505-272-5830; Fax: 505-272-8199; E-mail: sruby@unm.edu.
Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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