Characterization of fibroblast cytoplasmic proteins that bind to the 3' UTR of human catalase mRNA.
The excessive expression of catalase protein and its activity in cultured skin fibroblast from Zellweger Syndrome (ZS), a disorder of peroxisomal biogenesis, was found to be regulated at the translational level (J. Neurochem. 67: 2373-2378, 1996). Overall there is a considerable increase in the association of catalase mRNA with polysomes in ZS cell lines as compared to control indicating translational upregulation. To investigate the possibility that RNA-protein interactions are involved in the mediation of this increase in translation, the interaction between 3' untranslated region of human catalase mRNA and human fibroblast cytoplasmic proteins were investigated by RNA gel shift assay technique. Competition experiments demonstrated that all the 600 bases of 3' UTR (of human catalase gene) was required for efficient binding. Catalase RNA- protein interaction was sensitive to the altered redox state in these in vitro assays and this RNA-protein interaction could be enhanced by the addition of beta-mercaptoethanol in cytoplasm from control fibroblast but not in cytoplasm from ZS fibroblast. UV cross linked RNA-protein complexes on SDS polyacrylamide gel electrophoresis revealed the presence of at least four protein bands with approximate molecular masses of 38 kDa, 50 kDa, 66 kDa and 80 kDa. The potential role of these mRNA binding proteins in the regulation of catalase gene expression is discussed.
Ghosh
B
B
Department of Pediatrics, Medical University of South Carolina, Charleston 29425, USA.
Barbosa
E
E
Singh
I
I
D020413:3' Untranslated Regions / Q000378:metabolism* / Q000528:radiation effects; D002374:Catalase / Q000235:genetics*; D002460:Cell Line; D003593:Cytoplasm / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism; D006801:Humans; D010084:Oxidation-Reduction; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000378:metabolism* / Q000528:radiation effects; D016601:RNA-Binding Proteins / Q000378:metabolism* / Q000528:radiation effects; D012016:Reference Values; D012867:Skin / Q000378:metabolism*; D014158:Transcription, Genetic; D014466:Ultraviolet Rays; D015211:Zellweger Syndrome / Q000201:enzymology / Q000235:genetics
D020413:3' Untranslated Regions; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D002374:Catalase