Polysome-bound endonuclease PMR1 is targeted to stress granules via stress-specific binding to TIA-1.
The generalized process of mRNA decay involves deadenylation followed by release from translating polysomes, decapping, and exonuclease decay of the mRNA body. In contrast the mRNA endonuclease PMR1 forms a selective complex with its translating substrate mRNA, where it initiates decay by cleaving within the mRNA body. In stressed cells the phosphorylation of the alpha subunit of eukaryotic initiation factor 2 causes translating mRNAs to accumulate with stalled 48S subunits in large subcellular structures termed stress granules (SGs), wherein mRNAs undergo sorting for reinitiation, storage, or decay. Given the unique relationship between translation and PMR1-mediated mRNA decay, we examined the impact of stress-induced dissociation of polysomes on this process. Arsenite stress disrupts the polysome binding of PMR1 and its substrate mRNA but has no impact on the critical tyrosine phosphorylation of PMR1, its association with substrate mRNA, or its association with the functional approximately 680-kDa mRNP complex in which it normally resides on polysomes. We show that arsenite stress drives PMR1 into an RNase-resistant complex with TIA-1, and we identify a distinct domain in the N terminus of PMR1 that facilitates its interaction with TIA-1. Finally, we show that arsenite promotes the delayed association of PMR1 with SGs under conditions which cause tristetraprolin and butyrate response factor 1, proteins that facilitate exonucleolytic mRNA, to exit SGs.
Yang
Feng
F
Department of Molecular and Cellular Biochemistry, The Ohio State University, 1645 Neil Avenue, Columbus, OH 43210-1218, USA.
Peng
Yong
Y
Murray
Elizabeth L
EL
Otsuka
Yuichi
Y
Kedersha
Nancy
N
Schoenberg
Daniel R
DR
D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D003594:Cytoplasmic Granules / Q000378:metabolism*; D004722:Endoribonucleases / Q000235:genetics / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D039102:Poly(A)-Binding Proteins; D011132:Polyribosomes / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D013312:Stress, Physiological / Q000378:metabolism*; D000074019:T-Cell Intracellular Antigen-1; D014162:Transfection
D008565:Membrane Proteins; D039102:Poly(A)-Binding Proteins; D011993:Recombinant Fusion Proteins; D000074019:T-Cell Intracellular Antigen-1; C071558:TIA1 protein, human; D049452:Green Fluorescent Proteins; D004722:Endoribonucleases; C116181:polysomal RNase 1