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PMID: 1352851 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Separation of factors required for cleavage and polyadenylation of yeast pre-mRNA.

Molecular and cellular biology ·Vol. 12 ·No. 8 ·1992-08-00 ·Pages 3470-81

Chen J, Moore C

Abstract

Cleavage and polyadenylation of yeast precursor RNA require at least four functionally distinct factors (cleavage factor I [CF I], CF II, polyadenylation factor I [PF I], and poly(A) polymerase [PAP]) obtained from yeast whole cell extract. Cleavage of precursor occurs upon combination of the CF I and CF II fractions. The cleavage reaction proceeds in the absence of PAP or PF I. The cleavage factors exhibit low but detectable activity without exogenous ATP but are stimulated when this cofactor is included in the reaction. Cleavage by CF I and CF II is dependent on the presence of a (UA)6 sequence upstream of the GAL7 poly(A) site. The factors will also efficiently cleave precursor with the CYC1 poly(A) site. This RNA does not contain a UA repeat, and processing at this site is thought to be directed by a UAG...UAUGUA-type motif. Specific polyadenylation of a precleaved GAL7 RNA requires CF I, PF I, and a crude fraction containing PAP activity. The PAP fraction can be replaced by recombinant PAP, indicating that this enzyme is the only factor in this fraction needed for the reconstituted reaction. The poly(A) addition step is also dependent on the UA repeat. Since CF I is the only factor necessary for both cleavage and poly(A) addition, it is likely that this fraction contains a component which recognizes processing signals located upstream of the poly(A) site. The initial separation of processing factors in yeast cells suggests both interesting differences from and similarities to the mammalian system.

Related Genes
CYC1 GAL4
MeSH Terms
Base Sequence Chromosome Deletion DNA, Fungal/genetics Genes, Fungal Molecular Sequence Data Plasmids Poly A/genetics,metabolism Polynucleotide Adenylyltransferase/isolation & purification,metabolism RNA Precursors/genetics,metabolism RNA, Fungal/genetics,isolation & purification,metabolism RNA, Messenger/genetics,metabolism RNA-Binding Proteins/isolation & purification,metabolism Saccharomyces cerevisiae/genetics,metabolism Transcription, Genetic mRNA Cleavage and Polyadenylation Factors
Chemicals
DNA, Fungal RNA Precursors RNA, Fungal RNA, Messenger RNA-Binding Proteins mRNA Cleavage and Polyadenylation Factors Poly A Polynucleotide Adenylyltransferase
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Chen J
Department of Molecular Biology and Microbiology, Tufts University, Boston, Massachusetts 02111-1800.
Moore C
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.12.8.3470-3481.1992
Published
1992-08-00
Pages
3470-81
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC364596
Subset
IM
Grants
NIDDK NIH HHS · P30 DK34928 · United States
NIGMS NIH HHS · R01 GM41752-03 · United States
External Links
PubMed source DOI: 10.1128/mcb.12.8.3470-3481.1992 PMC Full Text View journal details
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