O-GlcNAcylation and Phosphorylation of β-Actin Serine199 in Diabetic Nephropathy.

The function of actin is regulated by various posttranslational modifications. We showed previously that in the kidney of the nonobese type 2 diabetes model Goto-Kakizaki (GK) rat, increased O-GlcNAcylation of β-actin protein is observed. It has also been reported that both O-GlcNAcylation and phosphorylation occur on serine199 of β-actin. However, their roles are not known. To elucidate their roles in diabetic nephropathy, we examined the rat kidney for changes in O-GlcNAcylation of serine199 (gS199)-actin and in phosphorylation of serine199 (pS199)-actin. Both gS199- and pS199-actin molecules had an apparent molecular weight of 40-kDa and were localized as non-filamentous actin both in the cytoplasm and in the nucleus. Compared with that in the normal kidney, the immunostaining intensity of gS199-actin increased in the podocytes of the glomeruli and in the proximal tubules of the diabetic kidney; whereas that of pS199-actin did not change in the podocytes but decreased in the proximal tubules. We confirmed that the same results could be observed in the glomeruli of the human diabetic kidney. In podocytes of glomeruli cultured in the presence of the O-GlcNAcase inhibitor Thiamet G, increased O-GlcNAcylation was accompanied by a concomitant decrease in the amount of filamentous-actin and in morphological changes. Our present results demonstrate that dysregulation of O-GlcNAcylation and phosphorylation of serine199 occurred in diabetes, which may contribute partially to the causes of the morphological changes in the glomeruli and tubules. gS199-actin and pS199-actin will thus be useful for the pathological evaluation of diabetic nephropathy.

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