Journal article

Publication year: 1987

Pages: 2415-2424

Journal: The EMBO Journal

Volume number: 6

Issue number: 8

ISSN: 0261-4189

eISSN: 1460-2075

URL: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC553648/

Publisher: EMBO Press

Abstract: 
We have studied the assembly, composition and structure of splicing complexes using biotin-avidin affinity chromatography and RNase protection assays. We find that U1, U2, U4, U5 and U6 snRNPs associate with the pre-mRNA and are in the mature, functional complex. Association of U1 snRNP with the pre-mRNA is rapid and ATP independent; binding of all other snRNPs occurs subsequently and is ATP dependent. Efficient binding of U1 and U2 snRNPs requires a 5' splice site or a 3' splice site/branch point region, respectively. Both sequence elements are required for efficient U4, U5 and U6 snRNP binding. Mutant RNA substrates containing only a 5' splice site or a 3' splice site/branch point region are assembled into 'partial' splicing complexes, which contain a subset of these five snRNPs. RNase protection experiments indicate that in contrast to U1 and U2 snRNPs, U4, U5 and U6 snRNPs do not contact the pre-mRNA. Based upon the time course of snRNP binding and the composition of sucrose gradient fractionated splicing complexes we suggest an assembly pathway proceeding from a 20S (U1 snRNP only) through a 40S (U1 and U2 snRNPs) to the functional 60S splicing complex (U1, U2, U4, U5 and U6 snRNPs).

Harvard Citation style: Bindereif, A. and Green, M. (1987) An ordered pathway of snRNP binding during mammalian pre-mRNA splicing complex assembly., The EMBO Journal, 6(8), pp. 2415-2424. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC553648/

APA Citation style: Bindereif, A., & Green, M. (1987). An ordered pathway of snRNP binding during mammalian pre-mRNA splicing complex assembly.. The EMBO Journal. 6(8), 2415-2424. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC553648/