An Ordered Pathway of SnRNP Binding During Mammalian Pre-mRNA Splicing Complex Assembly

Overview

Journal EMBO J

Specialties Biotechnology
Molecular Biology

Date 1987 Aug 1

PMID 2959470

Citations 122

Authors

A Bindereif

M R Green

Affiliations

Soon will be listed here.

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).

Citing Articles

An ATP-independent role for Prp16 in promoting aberrant splicing.

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The Role of Noncoding RNA Pseudouridylation in Nuclear Gene Expression Events.

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Identification of a 35S U4/U6.U5 tri-small nuclear ribonucleoprotein (tri-snRNP) complex intermediate in spliceosome assembly.

Chen Z, Gui B, Zhang Y, Xie G, Li W, Liu S J Biol Chem. 2017; 292(44):18113-18128.

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A composite double-/single-stranded RNA-binding region in protein Prp3 supports tri-snRNP stability and splicing.

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Insight into the mechanisms and functions of spliceosomal snRNA pseudouridylation.

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