Endonuclease-mediated mRNA decay requires tyrosine phosphorylation of polysomal ribonuclease 1 (PMR1) for the targeting and degradation of polyribosome-bound substrate mRNA.
PMR1 is an endonuclease that is activated by estrogen to degrade Xenopus albumin mRNA. A previous report showed that the functional unit of endonuclease-mediated mRNA decay is a approximately 680-kDa polysome-bound complex that contains both PMR1 and substrate mRNA. PMR1 contains two domains involved in endonuclease targeting to polysomes, an N-terminal domain that lies between residues 200 and 250, and a C-terminal domain that lies within the last 100 residues. Loss of either domain inactivated PMR1 targeting to polysomes and stabilized albumin mRNA. The current study identified a phosphorylated tyrosine residue within the C-terminal polysome-targeting domain and showed that this modification is required for PMR1-mediated mRNA decay. Changing this tyrosine to phenylalanine inactivated the targeting of PMR1 to polysomes, blocked binding of PMR1 to the functional complex containing its substrate mRNA, prevented the targeting of a green fluorescent protein fusion protein to this complex, and stabilized albumin mRNA to degradation by PMR1 in vivo. A general tyrosine kinase inhibitor inhibited the phosphorylation of PMR1, which in turn inhibited PMR1-catalyzed degradation of albumin mRNA. These results indicate that one or more tyrosine kinases functions as a regulator of endonuclease-mediated mRNA decay.
Yang
Feng
F
Department of Molecular and Cellular Biochemistry, Ohio State Biochemistry Program, Ohio State University, Colombus, OH 43210, USA.
Peng
Yong
Y
Schoenberg
Daniel R
DR
D000418:Albumins / Q000737:chemistry / Q000378:metabolism; D000818:Animals; D019556:COS Cells; D002499:Centrifugation, Density Gradient; D003593:Cytoplasm / Q000378:metabolism; D004720:Endonucleases / Q000378:metabolism*; D004722:Endoribonucleases / Q000737:chemistry* / Q000378:metabolism*; D017353:Gene Deletion; D007074:Immunoglobulin G / Q000737:chemistry; D047468:Immunoprecipitation; D008099:Liver / Q000378:metabolism; D008297:Male; D009154:Mutation; D010766:Phosphorylation; D010957:Plasmids / Q000378:metabolism; D011132:Polyribosomes / Q000737:chemistry; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors / Q000737:chemistry; D012333:RNA, Messenger / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012260:Ribonucleases / Q000378:metabolism; D012685:Sepharose / Q000494:pharmacology; D013395:Sucrose / Q000494:pharmacology; D014162:Transfection; D014443:Tyrosine / Q000737:chemistry*; D014981:Xenopus
D000418:Albumins; D007074:Immunoglobulin G; D012333:RNA, Messenger; D014443:Tyrosine; D013395:Sucrose; D012685:Sepharose; D011505:Protein-Tyrosine Kinases; D004720:Endonucleases; D004722:Endoribonucleases; D012260:Ribonucleases; C116181:polysomal RNase 1