The yeast POP2 gene encodes a nuclease involved in mRNA deadenylation.
The major mRNA degradation pathway involves deadenylation of the target molecule followed by decapping and, finally, 5'-->3' exonuclease digestion of the mRNA body. While yeast factors involved in the decapping and exonuclease degradation steps have been identified, the nature of the factor(s) involved in the deadenylation step remained elusive. Database searches for yeast proteins related to the mammalian deadenylase PARN identified the Pop2 protein (Pop2p) as a potential deadenylase. While Pop2p was previously identified as a factor affecting transcription, we identified a non-canonical RNase D sequence signature in its sequence. Analysis of the fate of a reporter mRNA in a pop2 mutant demonstrates that Pop2p is required for efficient mRNA degradation in vivo. Characterisation of mRNA degradation intermediates accumulating in this mutant supports the involvement of Pop2p in mRNA deadenylation in vivo. Similar phenotypes are observed in yeast strains lacking the Ccr4 protein, which is known to be associated with Pop2p. A recombinant Pop2p fragment encompassing the putative catalytic domain degrades poly(A) in vitro demonstrating that Pop2p is a nuclease. We also demonstrate that poly(A) is a better competitor than poly(G) or poly(C) of the Pop2p nuclease activity. Altogether, our study indicates that Pop2p is a nuclease subunit of the yeast deadenylase and suggests that Pop2p homologues in other species may have similar functions.
Daugeron
M C
MC
Centre de Génétique Moléculaire, CNRS, Avenue de la Terrasse, F-91198 Gif sur Yvette, France.
Mauxion
F
F
Séraphin
B
B
D000595:Amino Acid Sequence; D000818:Animals; D020134:Catalytic Domain; D019295:Computational Biology; D004722:Endoribonucleases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D017353:Gene Deletion; D005800:Genes, Fungal / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D011061:Poly A / Q000235:genetics; D017434:Protein Structure, Tertiary; D021122:Protein Subunits; D011506:Proteins; D020871:RNA Stability; D012331:RNA, Fungal / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D016914:Ribonuclease H / Q000378:metabolism; D043244:Ribonuclease III; D012260:Ribonucleases; D029701:Saccharomyces cerevisiae Proteins; D013379:Substrate Specificity; D014157:Transcription Factors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015003:Yeasts / Q000201:enzymology* / Q000235:genetics
C489780:CNOT8 protein, human; D005656:Fungal Proteins; D021122:Protein Subunits; D011506:Proteins; D012331:RNA, Fungal; D012333:RNA, Messenger; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; D011061:Poly A; C078657:CCR4 protein, S cerevisiae; D004722:Endoribonucleases; D012260:Ribonucleases; C079009:POP2 protein, S cerevisiae; D043244:Ribonuclease III; D016914:Ribonuclease H