PCF11 encodes a third protein component of yeast cleavage and polyadenylation factor I.
Cleavage and polyadenylation factor I (CF I) is one of four factors required in vitro for yeast pre-mRNA 3'-end processing. Two protein components of this factor, encoded by genes RNA14 and RNA15, have already been identified. We describe here another gene, PCF11 (for protein 1 of CF I), that genetically interacts with RNA14 and RNA15 and which presumably codes for a third protein component of CF I. This gene was isolated in a two-hybrid screening designed to identify proteins interacting with Rna14 and Rna15. PCF11 is an essential gene encoding for a protein of 626 amino acids having an apparent molecular mass of 70 kDa. Thermosensitive mutations in PCF11 are synergistically lethal with thermosensitive alleles of RNA14 and RNA15. The Pcf11-2 thermosensitive strain shows a shortening of the poly(A) tails and a strong decrease in the steady-state level of actin transcripts after a shift to the nonpermissive temperature as do the thermosensitive alleles of RNA14 and RNA15. Extracts from the pcf11-1 and pcf11-2 thermosensitive strains and the wild-type strain, when Pcf11 is neutralized by specific antibodies, are deficient in cleavage and polyadenylation. Moreover, fractions obtained by anion-exchange chromatography of extracts from the wild-type strain contain both Pcf11 and Rna15 in the same fractions, as shown by immunoblotting with a Pcf11-specific antibody.
Amrani
N
N
Centre de Génétique Moléculaire, CNRS UPR 9061, University of Paris VI (Pierre and Marie Curie), Gif sur Yvette, France.
Minet
M
M
Wyers
F
F
Dufour
M E
ME
Aggerbeck
L P
LP
Lacroute
F
F
D000595:Amino Acid Sequence; D003574:Cytochrome c Group / Q000235:genetics; D045304:Cytochromes c; D005656:Fungal Proteins / Q000235:genetics; D005800:Genes, Fungal / Q000235:genetics; D005804:Genes, Lethal; D008969:Molecular Sequence Data; D009154:Mutation; D009687:Nuclear Proteins / Q000235:genetics; D012323:RNA Processing, Post-Transcriptional / Q000235:genetics*; D012331:RNA, Fungal / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000235:genetics / Q000302:isolation & purification; D029701:Saccharomyces cerevisiae Proteins; D015003:Yeasts / Q000235:genetics*; D039221:mRNA Cleavage and Polyadenylation Factors
C014059:CYC1 protein, S cerevisiae; D003574:Cytochrome c Group; D005656:Fungal Proteins; D009687:Nuclear Proteins; C512637:PCF11 protein, S cerevisiae; D012331:RNA, Fungal; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D029701:Saccharomyces cerevisiae Proteins; D039221:mRNA Cleavage and Polyadenylation Factors; C512638:RNA14 protein, S cerevisiae; C512635:RNA15 protein, S cerevisiae; D045304:Cytochromes c