Identification of pp68 as the Tyrosine-phosphorylated Form of SYNCRIP/NSAP1. A cytoplasmic RNA-binding protein.
Recently we reported that osmotic shock increased the insulin-stimulated tyrosine phosphorylation of a 68-kDa RNA-binding protein in 3T3-L1 adipocytes (Hresko, R. C., and Mueckler, M. (2000) J. Biol. Chem. 275, 18114-18120). In this present study we have identified, by MALDI mass spectrometry, pp68 as the tyrosine-phosphorylated form of synaptotagmin-binding cytoplasmic RNA-interacting protein (SYNCRIP)/NSAP1, a newly discovered cytoplasmic RNA-binding protein. Both SYNCRIP and pp68 were enriched in free polysomes found in low density microsomes isolated from 3T3-L1 adipocytes. In vitro phosphorylation studies revealed that SYNCRIP, once extracted from low density microsomes, can be tyrosine phosphorylated using purified insulin receptor. Binding of RNA to SYNCRIP specifically inhibited its in vitro phosphorylation but had no effect on receptor autophosphorylation or on the ability of the receptor to phosphorylate a model substrate, RCM-lysozyme. These results raise the possibility that regulation of mRNA translation or stability by insulin may involve the phosphorylation of SYNCRIP.
Hresko
Richard C
RC
Department of Cell Biology and Physiology, Washington University School of Medicine, St. Louis, Missouri 63110, USA.
Mueckler
Mike
M
D016475:3T3 Cells; D000595:Amino Acid Sequence; D000818:Animals; D003593:Cytoplasm / Q000378:metabolism*; D034441:Heterogeneous-Nuclear Ribonucleoproteins; D051379:Mice; D008969:Molecular Sequence Data; D010750:Phosphoproteins / Q000737:chemistry / Q000378:metabolism*; D010766:Phosphorylation; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000737:chemistry / Q000378:metabolism*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013347:Subcellular Fractions / Q000378:metabolism; D014443:Tyrosine / Q000378:metabolism*
D034441:Heterogeneous-Nuclear Ribonucleoproteins; D010750:Phosphoproteins; D016601:RNA-Binding Proteins; C486049:Syncrip protein, mouse; D014443:Tyrosine; C410467:insulin receptor tyrosine kinase; D011505:Protein-Tyrosine Kinases