Evidence that poly(A) binding protein has an evolutionarily conserved function in facilitating mRNA biogenesis and export.
Eukaryotic poly(A) binding protein (PABP) is a ubiquitous, essential cellular factor with well-characterized roles in translational initiation and mRNA turnover. In addition, there exists genetic and biochemical evidence that PABP has an important nuclear function. Expression of PABP from Arabidopsis thaliana, PAB3, rescues an otherwise lethal phenotype of the yeast pab1Delta mutant, but it neither restores the poly(A) dependent stimulation of translation, nor protects the mRNA 5' cap from premature removal. In contrast, the plant PABP partially corrects the temporal lag that occurs prior to the entry of mRNA into the decay pathway in the yeast strains lacking Pab1p. Here, we examine the nature of this lag-correction function. We show that PABP (both PAB3 and the endogenous yeast Pab1p) act on the target mRNA via physically binding to it, to effect the lag correction. Furthermore, substituting PAB3 for the yeast Pab1p caused synthetic lethality with rna15-2 and gle2-1, alleles of the genes that encode a component of the nuclear pre-mRNA cleavage factor I, and a factor associated with the nuclear pore complex, respectively. PAB3 was present physically in the nucleus in the complemented yeast strain and was able to partially restore the poly(A) tail length control during polyadenylation in vitro, in a poly(A) nuclease (PAN)-dependent manner. Importantly, PAB3 in yeast also promoted the rate of entry of mRNA into the translated pool, rescued the conditional lethality, and alleviated the mRNA export defect of the nab2-1 mutant when overexpressed. We propose that eukaryotic PABPs have an evolutionarily conserved function in facilitating mRNA biogenesis and export.
Chekanova
Julia A
JA
Department of Biological Sciences, State University of New York at Albany, Albany, New York 12222, USA. dab@albany.edu
Belostotsky
Dmitry A
DA
D029681:Arabidopsis Proteins / Q000378:metabolism; D001692:Biological Transport; D019143:Evolution, Molecular; D017404:In Situ Hybridization, Fluorescence; D039102:Poly(A)-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D011485:Protein Binding; D012315:RNA Caps; D012331:RNA, Fungal / Q000096:biosynthesis* / Q000378:metabolism; D012333:RNA, Messenger / Q000096:biosynthesis* / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics
D029681:Arabidopsis Proteins; D039102:Poly(A)-Binding Proteins; D012315:RNA Caps; D012331:RNA, Fungal; D012333:RNA, Messenger; C450119:pab3 protein, Arabidopsis; C450101:poly(A)-binding protein 1, Arabidopsis