Nucleic Acids Research, 1990, Vol. 18, No. 5 1159-1166
© 1990
MOLECULAR BIOLOGY |
A cDNA from a mouse pancreatic ß cell encoding a putative transcription factor of the insulin gene
Department of Biochemistry, Welzmann Institute of Science 76100 Rehovot, Israel
*To whom correspondence should be addressed
Received December 5, 1989. Revised January 25, 1990. Accepted January 25, 1990.
Cell specific expression of the insulin gene is achieved through transcriptional mechanisms operating on multiple DNA sequence elements located in the 5' flanking region of the gene. Of particular importance in the rat insulin I gene are two closely similar 9 bp sequences (IEB1 and IEB2): mutation of either of these leads to 5 – 10 fold reduction in transcriptional activity. We have screened an expression cDNA library derived from mouse pancreatic endocrine ß cells with a radioactive DNA probe containing multiple copies of the IEB1 sequence. A cDNA clone (A1) isolated by this procedure encodes a protein which shows efficient binding to the IEB1 probe, but much weaker binding to either an unrelated DNA probe or to a probe bearing a single base pair insertion within the recognition sequence. DNA sequence analysis Indicates a protein belonging to the helix-loop-helix family of DNA-binding proteins. The ability of the protein encoded by clone A1 to recognize a number of wild type and mutant DNA sequences correlates closely with the ability of each sequence element to support transcription in vivo in the context of the insulin 5' flanking DNA. We conclude that the isolated cDNA may encode a transcription factor that participates in control of insulin gene expression.
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