The cytoskeleton-associated Ena/VASP proteins are unanticipated partners of the PMR1 mRNA endonuclease.
The PMR1 mRNA endonuclease catalyzes the selective decay of a limited number of mRNAs. It participates in multiple complexes, including one containing c-Src, its activating kinase, and one containing its substrate mRNA. This study used tandem affinity purification (TAP) chromatography to identify proteins in HeLa cell S100 associated with the mature 60-kDa form of Xenopus PMR1 (xPMR60). Unexpectedly, this identified a number of cytoskeleton-associated proteins, most notably the Ena family proteins mammalian Enabled (Mena) and vasodilator-stimulated phosphoprotein (VASP). These are regulators of actin dynamics that distribute throughout the cytoplasm and concentrate along the leading edge of the cell. xPMR60 interacts with Mena and VASP in vivo, overexpression of Mena has no impact on mRNA decay, and Mena and VASP are recovered together with xPMR60 in each of the major complexes of PMR1-mRNA decay. In a wound-healing experiment induced expression of active xPMR60 in stably transfected cells resulted in a twofold increase in cell motility compared with uninduced cells or cells expressing inactive xPMR60 degrees . Under these conditions xPMR60 colocalizes with VASP along one edge of the cell.
Peng
Yong
Y
Department of Molecular and Cellular Biochemistry, The Ohio State University, Columbus, 43210, USA.
Murray
Elizabeth L
EL
Sarkar
Madhubanti
M
Liu
Xiaoqiang
X
Schoenberg
Daniel R
DR
D000199:Actins / Q000378:metabolism; D000818:Animals; D019556:COS Cells; D015815:Cell Adhesion Molecules / Q000302:isolation & purification / Q000378:metabolism*; D002465:Cell Movement; D002522:Chlorocebus aethiops; D002846:Chromatography, Affinity; D004722:Endoribonucleases / Q000235:genetics*; D006367:HeLa Cells; D006801:Humans; D008840:Microfilament Proteins / Q000302:isolation & purification / Q000378:metabolism*; D010750:Phosphoproteins / Q000302:isolation & purification / Q000378:metabolism*; D020871:RNA Stability; D014162:Transfection; D029867:Xenopus Proteins / Q000235:genetics*; D000098662:Vasodilator-Stimulated Phosphoprotein
D000199:Actins; D015815:Cell Adhesion Molecules; D008840:Microfilament Proteins; D010750:Phosphoproteins; D029867:Xenopus Proteins; D000098662:Vasodilator-Stimulated Phosphoprotein; D004722:Endoribonucleases; C539513:PMR-1 polysomal ribonuclease 1, Xenopus laevis