ADP-ribosylation of the Mr 83,000 stress-inducible and glucose-regulated protein in avian and mammalian cells: Modulation by heat shock and glucose starvation
ADP-ribosylation of proteins was analyzed by in vivo labeling of cells with [3H]adenosine, followed by separation of their protein components by two-dimensional isoelectric focusing/NaDodSO4 polyacrylamide gel electrophoresis. We show here that in several cell types of avian and mammalian origin the major [3H]adenosine acceptor in vivo is a polypeptide with a Mr of 83,000 and isoelectric point of ≈5.3. This polypeptide is identical to one of the stress-inducible and glucose-regulated proteins (here called SP83) previously described in avian and mammalian cells. Snake venom phosphodiesterase digestion of purified 3H-labeled SP83 releases 5'-AMP and a minor fraction of 2'-(5' '}-phosphoribosyl)-5'-AMP. In vitro labeling with [32P]NAD+ of total cell lysates made in the presence of non-ionic detergents also results in incorporation of radioactivity into SP83. Both of these results strongly suggest that the modification is an ADP-ribosylation. Heat shock and glucose starvation of cells induce a rapid and extensive decrease in the incorporation of ADP-ribose into SP83, suggesting that ADP-ribosylation may be important for the regulation of the function of this protein.
Additional Information© 1983 by the National Academy of Sciences. Communicated by James F. Bonner, April 26, 1983. We thank Drs. I. Blikstad and B. L. Granger for many valuable comments on this work. This work was supported by grants from the National Institutes of Health (GM 06965), National Science Foundation, and Muscular Dystrophy Association of America. L.C. was a recipient of a European Molecular Biology Organization Long-Term Fellowship during the course of this investigation. E.L. is a recipient of a Research Career Development Award from the National Institutes of Health. The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.
Published - CARpnas83.pdf