Control of the ATP synthase beta subunit expression by RNA-binding proteins TIA-1, TIAR, and HuR.
The beta-subunit of the mitochondrial H+-ATP synthase (beta-F1-ATPase) catalyzes the rate-limiting step of ATP formation in eukaryotic cells. Here, we examined the post-transcriptional regulation of human beta-F1-ATPase mediated by the 3'-untranslated region of the mRNA (beta-3'-UTR). Biochemical analysis revealed that the adenosine/uridine (AU)-rich element-binding proteins TIA-1 (T-cell intracellular antigen-1), TIAR (TIA-1-related protein), and HuR (Hu antigen R) interact with the beta-F1-ATPase mRNA through an AU-rich sequence located to the 3'-UTR. Mouse embryonic fibroblasts (MEFs) knocked-out for TIA-1 or RNA interference (RNAi)-mediated knockdown of endogenous TIA-1, TIAR, or HuR in HeLa cells resulted in a decrease in beta-F1-ATPase protein expression. The expression of GFP from a chimeric reporter containing human beta-3'-UTR was also abolished in HeLa cells depleted of TIA-1, TIAR, or HuR. MEFs knocked-in for TIA-1 or the overexpression of RNAi-resistant TIA-1, TIAR, or HuR proteins in the RNAi-treated HeLa cells significantly restored the levels of the expression of both endogenous mouse beta-F1-ATPase protein or recombinant GFP.
Izquierdo
José M
JM
Centro de Biología Molecular Severo Ochoa, Universidad Autónoma de Madrid (UAM-CSIC), Facultad de Ciencias, Módulo C-V, Lab-230, Cantoblanco, DP 28049, Madrid, Spain.
D020413:3' Untranslated Regions / Q000502:physiology; D000818:Animals; D000954:Antigens, Surface / Q000502:physiology*; D001483:Base Sequence; D051959:ELAV Proteins; D000067780:ELAV-Like Protein 1; D005786:Gene Expression Regulation; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D018345:Mice, Knockout; D025261:Mitochondrial Proton-Translocating ATPases / Q000096:biosynthesis* / Q000235:genetics*; D016601:RNA-Binding Proteins / Q000502:physiology*; D016415:Sequence Alignment
D020413:3' Untranslated Regions; C506526:ATP5F1B protein, human; D000954:Antigens, Surface; D051959:ELAV Proteins; D000067780:ELAV-Like Protein 1; C098770:ELAVL1 protein, human; D016601:RNA-Binding Proteins; C076751:TIAL1 protein, human; D025261:Mitochondrial Proton-Translocating ATPases