Functional analysis of the human CDC5L complex and identification of its components by mass spectrometry
- 1 December 2000
- journal article
- Published by Springer Science and Business Media LLC in The EMBO Journal
- Vol. 19 (23), 6569-6581
- https://doi.org/10.1093/emboj/19.23.6569
Abstract
Recently, we identified proteins that co‐purify with the human spliceosome using mass spectrometry. One of the identified proteins, CDC5L, corresponds to the human homologue of the Schizosaccharomyces pombe CDC5+ gene product. Here we show that CDC5L is part of a larger multiprotein complex in HeLa nuclear extract that incorporates into the spliceosome in an ATP‐dependent step. We also show that this complex is required for the second catalytic step of pre‐mRNA splicing. Immunodepletion of the CDC5L complex from HeLa nuclear extract inhibits the formation of pre‐mRNA splicing products in vitro but does not prevent spliceosome assembly. The first catalytic step of pre‐mRNA splicing is less affected by immunodepleting the complex. The purified CDC5L complex in HeLa nuclear extract restores pre‐mRNA splicing activity when added to extracts that have been immunodepleted using anti‐CDC5L antibodies. Using mass spectrometry and database searches, the major protein components of the CDC5L complex have been identified. This work reports a first purification and characterization of a functional, human non‐snRNA spliceosome subunit containing CDC5L and at least five additional protein factors.Keywords
This publication has 57 references indexed in Scilit:
- NIPP1-mediated Interaction of Protein Phosphatase-1 with CDC5L, a Regulator of Pre-mRNA Splicing and Mitotic EntryJournal of Biological Chemistry, 2000
- RBMY, a probable human spermatogenesis factor, and other hnRNP G proteins interact with Tra2beta and affect splicingHuman Molecular Genetics, 2000
- Rearrangement of the HumanCDC5LGene by a t(6;19)(p21;q13.1) in a Patient with Multicystic Renal DysplasiaGenomics, 1998
- Structural basis for the recognition of regulatory subunits by the catalytic subunit of protein phosphatase 1The EMBO Journal, 1997
- Interaction of protein phosphatase type 1 with a splicing factorFEBS Letters, 1996
- Correspondence Between a Mammalian Spliceosome Component and an Essential Yeast Splicing FactorScience, 1993
- Intracellular distribution of the U1A protein depends on active transport and nuclear binding to U1 snRNA.The Journal of cell biology, 1992
- A mutational analysis of spliceosome assembly: evidence for splice site collaboration during spliceosome formation.Genes & Development, 1987
- Interactions between small nuclear ribonucleoprotein particles in formation of spliceosomesCell, 1987
- Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4Nature, 1970