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PMID: 11060040 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Human pre-mRNA cleavage factor II(m) contains homologs of yeast proteins and bridges two other cleavage factors.

The EMBO journal ·Vol. 19 ·No. 21 ·2000-11-01 ·Pages 5895-904

de Vries H, Rüegsegger U, Hübner W, Friedlein A, Langen H, Keller W

Abstract

Six different protein factors are required in vitro for 3' end formation of mammalian pre-mRNAs by endonucleolytic cleavage and polyadenylation. Five of the factors have been purified and most of their components cloned, but cleavage factor II(m) (CF II(m)) remained uncharacterized. We have purified CF II(m) from HeLa cell nuclear extract by several chromatographic steps. During purification, CF II(m) activity separated into two components, one essential (CF IIA(m)) and one stimulatory (CF IIB(m)) for the cleavage reaction. CF IIA(m) fractions contain the human homologs of two yeast 3' end processing factors, Pcf11p and Clp1p, as well as cleavage factor I(m) (CF I(m)) and several splicing and transcription factors. We report the cloning of hClp1 and show that it is a genuine subunit of CF IIA(m). Antibodies directed against hClp1 deplete cleavage activity, but not polyadenylation activity from HeLa cell nuclear extract. hClp1 interacts with CF I(m) and the cleavage and polyadenylation specificity factor CPSF, suggesting that it bridges these two 3' end processing factors within the cleavage complex.

MeSH Terms
Amino Acid Sequence Base Sequence Conserved Sequence DNA Primers/genetics Evolution, Molecular Fungal Proteins/chemistry,genetics HeLa Cells Humans Molecular Sequence Data Protein Subunits RNA Precursors/metabolism RNA Processing, Post-Transcriptional RNA Splicing RNA, Fungal/metabolism RNA-Binding Proteins/chemistry,genetics,metabolism Saccharomyces cerevisiae/genetics,metabolism Sequence Homology, Amino Acid Transcription, Genetic mRNA Cleavage and Polyadenylation Factors
Chemicals
DNA Primers Fungal Proteins Protein Subunits RNA Precursors RNA, Fungal RNA-Binding Proteins mRNA Cleavage and Polyadenylation Factors
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
de Vries H
Department of Cell Biology, Biozentrum, University of Basel, Klingelbergstrasse 70, CH-4056 Basel, Germany.
Rüegsegger U
Hübner W
Friedlein A
Langen H
Keller W
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1093/emboj/19.21.5895
Published
2000-11-01
Pages
5895-904
Language
English
Region
England
NLM ID
8208664
PMCID
PMC305781
Subset
IM
External Links
PubMed source DOI: 10.1093/emboj/19.21.5895 PMC Full Text View journal details
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