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PMID: 1383687 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

The spliceosome assembly pathway in mammalian extracts.

Molecular and cellular biology ·Vol. 12 ·No. 10 ·1992-10-00 ·Pages 4279-87

Jamison SF, Crow A, Garcia-Blanco MA

Abstract

A mammalian splicing commitment complex was functionally defined by using a template commitment assay. This complex was partially purified and shown to be a required intermediate for complex A formation. The productive formation of this commitment complex required both splice sites and the polypyrimidine tract. U1 small nuclear ribonucleoprotein (snRNP) was the only spliceosomal U snRNP required for this formation. A protein factor, very likely U2AF, is probably involved in the formation of the splicing commitment complex. From the kinetics of appearance of complex A and complex B, it was previously postulated that complex A represents a functional intermediate in spliceosome assembly. Complex A was partially purified and shown to be a required intermediate for complex B (spliceosome) formation. Thus, a spliceosome pathway is for the first time supported by direct biochemical evidence: RNA+U1 snRNP+?U2 auxiliary factor+?Y----CC+U2 snRNP+Z----A+U4/6,5 snRNPs+ beta----B.

MeSH Terms
Base Sequence Cell Nucleus/metabolism Centrifugation, Density Gradient DNA HeLa Cells Humans Introns Molecular Sequence Data Nuclear Proteins/metabolism RNA RNA Splicing Ribonucleoproteins, Small Nuclear/metabolism
Chemicals
Nuclear Proteins Ribonucleoproteins, Small Nuclear RNA DNA
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Jamison S F
Department of Microbiology and Immunology, Duke University Medical Center, Durham, North Carolina 27710.
Crow A
Garcia-Blanco M A
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41 references, click to expand
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.12.10.4279-4287.1992
Published
1992-10-00
Pages
4279-87
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC360351
Subset
IM
External Links
PubMed source DOI: 10.1128/mcb.12.10.4279-4287.1992 PMC Full Text View journal details
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