Characterization of the human CUTA isoform2 present in the stably transfected HeLa cells
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CUTA, Homo sapiens divalent cation tolerance homolog, has been implicated in anchoring of acetylcholinesterase in neuronal cell membranes. However, a protein highly homologous to CUTA in Rattus norvegicus is structurally similar to the signal transduction protein PII, and this similarity suggests an intriguing role of CUTA in signal transduction. Recent researches indicated that CUTA was one of the 35 key genes responsible for lactation in mammary gland development. However, the physiological role of CUTA is still unclear, so more information of this gene is needed. In this study, the expression profile of CUTA gene in human tissues was examined, and our research revealed that CUTA gene was constitutively expressed in all of the 18 tissues tested. As reported, CUTA gene has five variant transcripts encoding three isoforms with different N terminals. CUTA isoform2 is encoded by three of the five variant transcripts as the common part of the three isoforms. So CUTA isoform2 was chose as representative to characterize the CUTA protein. We constructed a HeLa cell line stably transfected with the encoding sequence of CUTA isoform2 for further study. The subcellular location and oligomeric structure of the CUTA isoform2 was analyzed in the stable cell lines. It was found that the CUTA isoform2 was mainly located in mitochondria as a new potential mitochondrial protein. Furthermore, CUTA isoform2 formed trimers in cell lysate with the possible occurrence of heteropolymers. These findings would be helpful to the further study on the specific function of CUTA protein.
KeywordsCUTA Stable transfection Mitochondria Cross-linking Trimeric structure
This work was supported by the National 973 program of China, 863 projects of China and the National Natural Science foundation of China (30024001).
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