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Journal Article
Research Support, Non-U.S. Gov't
Transcriptional regulation of insulin receptor gene promoter in rat hepatocytes.
Biochemical and Biophysical Research Communications 2001 Februrary 10
To investigate the DNA regulatory sequences -618 to -593 (initiator ATG is +1) of the insulin receptor (IR) promoter required for IR gene expression, primary cultured hepatocytes of rats were transfected with plasmids containing the 5'-flanking sequences of the rat IR gene fused to the luciferase gene. When three copies of the nucleotides -618 to -593 of the IR promoter were transfected, the reporter activity was significantly increased in the presence of glucose and more increased in the presence of glucose/insulin. The glucose/insulin stimulation was inhibited by the addition of polyunsaturated fatty acids. These results were similar to those found earlier for the transcriptions of the fatty acid synthase, FAS(-57/-35), ATP citrate-lyase, ACL(-64/-41), and leptin(-101/-83) genes, which promoter sequences have a high similarity to IR(-618/-593). However, similarly to the leptin gene, the IR gene was more responsive to glucose stimulation than the FAS and ACL genes. Mutation of either one of the Sp1 binding sites (-618/-593) did not significantly affect the transcription, whereas mutation of three or four Sp1 binding sites resulted in a loss of responsiveness to glucose/insulin. Gel mobility shift assays revealed that nuclear factor(s) from rat liver specifically formed complexes with the sequence of IR(-618/-593). By antibody supershift assays, the transcription factors Sp1and Sp3 were found to bind with the IR(-618/-593). The IR, leptin, ACL and FAS genes contain common DNA-sequences responsible for the glucose/insulin-stimulation, suggesting that these genes are similarly regulated.
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