PAB1 self-association precludes its binding to poly(A), thereby accelerating CCR4 deadenylation in vivo.
The mRNA deadenylation process, catalyzed by the CCR4 deadenylase, is known to be the major factor controlling mRNA decay rates in Saccharomyces cerevisiae. We have identified the proline-rich region and RRM1 domains of poly(A) binding protein (PAB1) as necessary for CCR4 deadenylation. Deletion of either of these regions but not other regions of PAB1 significantly reduced PAB1-PAB1 protein interactions, suggesting that PAB1 oligomerization is a required step for deadenylation. Moreover, defects in these two regions inhibited the formation of a novel, circular monomeric PAB1 species that forms in the absence of poly(A). Removal of the PAB1 RRM3 domain, which promoted PAB1 oligomerization and circularization, correspondingly accelerated CCR4 deadenylation. Circular PAB1 was unable to bind poly(A), and PAB1 multimers were severely deficient or unable to bind poly(A), implicating the PAB1 RNA binding surface as critical in making contacts that allow PAB1 self-association. These results support the model that the control of CCR4 deadenylation in vivo occurs in part through the removal of PAB1 from the poly(A) tail following its self-association into multimers and/or a circular species. Known alterations in the P domains of different PAB proteins and factors and conditions that affect PAB1 self-association would, therefore, be expected to be critical to controlling mRNA turnover in the cell.
Yao
Gang
G
Department of Biochemistry and Molecular Biology, Rudman Hall, University of New Hampshire, Durham, NH 03824, USA.
Chiang
Yueh-Chin
YC
Zhang
Chongxu
C
Lee
Darren J
DJ
Laue
Thomas M
TM
Denis
Clyde L
CL
D008958:Models, Molecular; D039102:Poly(A)-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D026723:Polyadenylation; D011392:Proline / Q000378:metabolism; D011487:Protein Conformation; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*; D012260:Ribonucleases / Q000235:genetics / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
D039102:Poly(A)-Binding Proteins; D020033:Protein Isoforms; D012333:RNA, Messenger; D029701:Saccharomyces cerevisiae Proteins; C450100:pab1 protein, S cerevisiae; D011392:Proline; C078657:CCR4 protein, S cerevisiae; D012260:Ribonucleases