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. 1992 Jun 15;89(12):5482–5486. doi: 10.1073/pnas.89.12.5482

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

S F Jamison 1, M A Garcia-Blanco 1
PMCID: PMC49316  PMID: 1535158

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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Selected References

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