title: PUF3 acceleration of deadenylation in vivo can operate independently of CCR4 activity, possibly involving effects on the PAB1-mRNP structure.
abstract: The evolutionarily conserved PUF proteins stimulate CCR4 mRNA deadenylation through binding to 3' untranslated region sequences of specific mRNA. We have investigated the mechanisms by which PUF3 in Saccharomyces cerevisiae accelerates deadenylation of the COX17 mRNA. PUF3 was shown to affect PAN2 deadenylation of the COX17 mRNA independent of the presence of CCR4, suggesting that PUF3 acts through a general mechanism to affect deadenylation. Similarly, eIF4E, the cap-binding translation initiation factor known to control CCR4 deadenylation, was shown to affect PAN2 activity in vivo. PUF3 was found to be required for eIF4E effects on COX17 deadenylation. Both eIF4E and PUF3 effects on deadenylation were shown, in turn, to necessitate a functional poly(A)-binding protein (PAB1) in which removal of the RRM1 (RNA recognition motif 1) domain of PAB1 blocked both their effects on deadenylation. While removal of the proline-rich region (P domain) of PAB1 substantially reduces CCR4 deadenylation at non-PUF3-controlled mRNA and correspondingly blocked eIF4E effects on deadenylation, PUF3 essentially bypassed this P domain requirement. These results indicate that the PAB1-mRNP structure is critical for PUF3 action. We also found that multiple components of the CCR4-NOT deadenylase complex, but not PAN2, interacted with PUF3. PUF3 appears, therefore, both to act independently of CCR4 activity, possibly through effects on PAB1-mRNP structure, and to be capable of retaining the CCR4-NOT complex.
authors.affiliation: Department of Biochemistry and Molecular Biology, Rudman Hall, University of New Hampshire, Durham, NH 03824, USA.
authors.forename: Darren
authors.initials: D
authors.lastname: Lee
authors.forename: Takbum
authors.initials: T
authors.lastname: Ohn
authors.forename: Yueh-Chin
authors.initials: YC
authors.lastname: Chiang
authors.forename: Gloria
authors.initials: G
authors.lastname: Quigley
authors.forename: Gang
authors.initials: G
authors.lastname: Yao
authors.forename: Yuting
authors.initials: Y
authors.lastname: Liu
authors.forename: Clyde L
authors.initials: CL
authors.lastname: Denis
mesh_terms: D027682:Cation Transport Proteins / Q000235:genetics / Q000378:metabolism; D000079922:Copper Transport Proteins; D039561:Eukaryotic Initiation Factor-4E / Q000235:genetics / Q000378:metabolism; D005095:Exoribonucleases / Q000235:genetics / Q000378:metabolism; D005800:Genes, Fungal; D007700:Kinetics; D018832:Molecular Chaperones / Q000235:genetics / Q000378:metabolism; D046912:Multiprotein Complexes; D039102:Poly(A)-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D012331:RNA, Fungal / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012260:Ribonucleases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012261:Ribonucleoproteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D027682:Cation Transport Proteins; D000079922:Copper Transport Proteins; D039561:Eukaryotic Initiation Factor-4E; D005095:Exoribonucleases; D018832:Molecular Chaperones; D046912:Multiprotein Complexes; D039102:Poly(A)-Binding Proteins; D012331:RNA, Fungal; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D011993:Recombinant Fusion Proteins; D012260:Ribonucleases; D012261:Ribonucleoproteins; D029701:Saccharomyces cerevisiae Proteins; C100359:COX17 protein, S cerevisiae; C510155:PUF3 protein, S cerevisiae; C026452:messenger ribonucleoprotein; C450100:pab1 protein, S cerevisiae; C078657:CCR4 protein, S cerevisiae