Poly(A)-binding-protein-mediated regulation of hDcp2 decapping in vitro.
Regulation of mRNA decapping is a critical determinant for gene expression. We demonstrate that the poly(A) tail-mediated regulation of mRNA decapping observed in humans can be recapitulated in vitro by the cytoplasmic poly(A)-binding protein PABP through a direct and specific binding to the 5' end of capped mRNA. The specific association of PABP with the cap occurred only within the context of the RNA whereby a cap attached to an RNA moiety served as the high-affinity substrate but not the cap structure or RNA alone. Binding of PABP to the RNA 5' end required the presence of the cap and was accentuated by the N7 methyl moiety of the cap. Interestingly, conditions that enhanced hDcp2 decapping activity reduced the affinity of PABP for cap association and consequently its ability to inhibit decapping, suggestive of a regulated association of PABP with the cap. These observations reveal a novel direct involvement of human PABP in the stabilization of mRNA by protecting the 5' end from decapping.
Khanna
Richie
R
Department of Cell Biology and Neuroscience, Rutgers University, Piscataway, NJ, USA.
Kiledjian
Megerditch
M
D017931:DNA Primers; D024202:Electrophoretic Mobility Shift Assay; D004722:Endoribonucleases / Q000378:metabolism* / Q000528:radiation effects; D006801:Humans; D047468:Immunoprecipitation; D010957:Plasmids / Q000235:genetics; D039102:Poly(A)-Binding Proteins / Q000378:metabolism*; D012315:RNA Caps / Q000378:metabolism*; D020871:RNA Stability / Q000502:physiology*; D014466:Ultraviolet Rays
D017931:DNA Primers; D039102:Poly(A)-Binding Proteins; D012315:RNA Caps; D004722:Endoribonucleases; C467723:DCP2 protein, human