Endonuclease-mediated mRNA decay involves the selective targeting of PMR1 to polyribosome-bound substrate mRNA.
PMR1 is a polysome-associated mRNA endonuclease that initiates the destabilization of albumin mRNA. The current study examined whether endonuclease-mediated mRNA decay involved the selective binding of PMR1 to substrate mRNA on polysomes. PMR1 is uniformly distributed throughout the cytoplasm on polysomes and in lighter complexes and does not colocalize in cytoplasmic foci with Dcp1. Deletion mutagenesis identified polysome-targeting domains in the N and C termini of PMR1, either of which could target GFP to polysomes. Selectivity in targeting to polysome-bound substrate mRNP was determined by testing the ability of full-length PMR1 or protein lacking targeting domains to recover albumin and luciferase mRNA from dissociated polysomes. Only PMR1 bearing intact polysome-targeting domains selectively recovered albumin mRNA, and polysome targeting of both protein and substrate was required for the efficient degradation of albumin mRNA. Thus, endonuclease-mediated mRNA decay occurs on a polysome-bound complex containing PMR1 and its substrate mRNA.
Yang
Feng
F
Department of Molecular and Cellular Biochemistry, The Ohio State Biochemistry Program, The Comprehensive Cancer Center, The Ohio State University, Columbus, OH 43210, USA.
Schoenberg
Daniel R
DR
D018528:ATP-Binding Cassette Transporters / Q000235:genetics / Q000378:metabolism*; D000418:Albumins / Q000235:genetics / Q000378:metabolism; D000818:Animals; D019556:COS Cells; D004720:Endonucleases / Q000235:genetics / Q000378:metabolism*; D004722:Endoribonucleases / Q000235:genetics; D008156:Luciferases / Q000235:genetics; D011132:Polyribosomes / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding / Q000235:genetics; D017434:Protein Structure, Tertiary / Q000235:genetics; D021381:Protein Transport / Q000235:genetics / Q000502:physiology; D020871:RNA Stability / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000235:genetics; D012261:Ribonucleoproteins / Q000378:metabolism
D018528:ATP-Binding Cassette Transporters; D000418:Albumins; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D012261:Ribonucleoproteins; C026452:messenger ribonucleoprotein; D008156:Luciferases; D004720:Endonucleases; D004722:Endoribonucleases