Seasonal variation of macrobenthic infauna in the Johor Strait, Singapore

10.1007/s10452-004-7111-2Aquatic Ecology391-2107-111AQEC

Y-box protein-1 controls transforming growth factor-beta1 translation in proximal tubular cells

Transforming growth factor-1 (TGF-1) mRNA has low basal translational efficiency in proximal tubule cells; however, its translation is stimulated by profibrotic cytokines. We studied the role of the multifunctional Y-box protein-1 (YB-1) in regulating proximal tubule cell TGF-1 translation. Using RNA-electrophoretic mobility shift assays and ultraviolet crosslinking, we found two protein complexes of 50 and 100 kDa, which bound to the TGF-1 mRNA 5'-untranslated region. Supershift studies using antibodies to YB-1 showed that both sites contained YB-1 as did studies with recombinant YB-1, which demonstrated that it was sufficient to form both complexes. RNA competition experiments confirmed YB-1 binding to the two predicted binding sites; one with high affinity and the other with lower affinity. Strong basal YB-1 association with TGF-1 mRNA was found in proximal tubule cells, which decreased when platelet-derived growth factor was used to activate TGF-1 translation. In contrast, knockdown of proximal tubule cell YB-1 expression abrogated TGF-1 synthesis. Our results suggest that TGF-1 translation in proximal tubule cells requires YB-1 binding to a high-affinity site in the 5'-untranslated region of its mRNA; however, binding to a low-affinity site inhibits basal translation