An ATP-independent U2 small nuclear ribonucleoprotein particle/precursor mRNA complex requires both splice sites and the polypyrimidine tract

Sharon F. Jamison, Mariano Garcia-Blanco

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

A complex is formed upon incubation of a precursor mRNA (pre-mRNA) with HeLa cell nuclear extract in the absence of added ATP (-ATP complex). Pre-mRNAs with mutations in the 5′ splice site, the 3′ splice site, or the polypyrimidine tract did not form this complex. Once formed, the -ATP complex was stable to competition by excess pre-mRNA. The complex was shown to contain the U2 small nuclear ribonucleoprotein particle (snRNP) and was distinct from the previously described U2 snRNP/pre-mRNA complex, the prespliceosome. These complexes have different electrophoretic mobilities, ATP requirements, and sensitivities to mutations of the 5' splice site. Although U1 snRNP was not found in the -ATP complex, a requirement for the U1 snRNP was suggested by immunodepletion experiments. The possible implications for the study of spliceosome formation are discussed.

Original languageEnglish (US)
Pages (from-to)5482-5486
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume89
Issue number12
StatePublished - 1992
Externally publishedYes

Fingerprint

U2 Small Nuclear Ribonucleoproteins
Small Nuclear Ribonucleoproteins
RNA Precursors
Adenosine Triphosphate
RNA Splice Sites
U1 Small Nuclear Ribonucleoproteins
Spliceosomes
Mutation
Cell Extracts
HeLa Cells

Keywords

  • Precursor mRNA splicing
  • Prespliceosome
  • Spliceosome

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

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abstract = "A complex is formed upon incubation of a precursor mRNA (pre-mRNA) with HeLa cell nuclear extract in the absence of added ATP (-ATP complex). Pre-mRNAs with mutations in the 5′ splice site, the 3′ splice site, or the polypyrimidine tract did not form this complex. Once formed, the -ATP complex was stable to competition by excess pre-mRNA. The complex was shown to contain the U2 small nuclear ribonucleoprotein particle (snRNP) and was distinct from the previously described U2 snRNP/pre-mRNA complex, the prespliceosome. These complexes have different electrophoretic mobilities, ATP requirements, and sensitivities to mutations of the 5' splice site. Although U1 snRNP was not found in the -ATP complex, a requirement for the U1 snRNP was suggested by immunodepletion experiments. The possible implications for the study of spliceosome formation are discussed.",
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T1 - An ATP-independent U2 small nuclear ribonucleoprotein particle/precursor mRNA complex requires both splice sites and the polypyrimidine tract

AU - Jamison, Sharon F.

AU - Garcia-Blanco, Mariano

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N2 - A complex is formed upon incubation of a precursor mRNA (pre-mRNA) with HeLa cell nuclear extract in the absence of added ATP (-ATP complex). Pre-mRNAs with mutations in the 5′ splice site, the 3′ splice site, or the polypyrimidine tract did not form this complex. Once formed, the -ATP complex was stable to competition by excess pre-mRNA. The complex was shown to contain the U2 small nuclear ribonucleoprotein particle (snRNP) and was distinct from the previously described U2 snRNP/pre-mRNA complex, the prespliceosome. These complexes have different electrophoretic mobilities, ATP requirements, and sensitivities to mutations of the 5' splice site. Although U1 snRNP was not found in the -ATP complex, a requirement for the U1 snRNP was suggested by immunodepletion experiments. The possible implications for the study of spliceosome formation are discussed.

AB - A complex is formed upon incubation of a precursor mRNA (pre-mRNA) with HeLa cell nuclear extract in the absence of added ATP (-ATP complex). Pre-mRNAs with mutations in the 5′ splice site, the 3′ splice site, or the polypyrimidine tract did not form this complex. Once formed, the -ATP complex was stable to competition by excess pre-mRNA. The complex was shown to contain the U2 small nuclear ribonucleoprotein particle (snRNP) and was distinct from the previously described U2 snRNP/pre-mRNA complex, the prespliceosome. These complexes have different electrophoretic mobilities, ATP requirements, and sensitivities to mutations of the 5' splice site. Although U1 snRNP was not found in the -ATP complex, a requirement for the U1 snRNP was suggested by immunodepletion experiments. The possible implications for the study of spliceosome formation are discussed.

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