PMID: 9421492Feb 28, 1998Paper

Overlapping Sp1 and AP2 binding sites in a promoter element of the lens-specific MIP gene

Nucleic Acids Research
C Ohtaka-MaruyamaA B Chepelinsky

Abstract

The MIP gene, the founder of the MIP family of channel proteins, is specifically expressed in fiber cells of the ocular lens and expression is regulated temporally and spatially during development. We previously found that a DNA fragment containing 253 bp of 5'-flanking sequence and 42 bp of exon 1 of the human MIP gene contains regulatory elements responsible for lens-specific expression of the MIP gene. In this report we have analyzed the function of overlapping Sp1 and AP2 binding sites present in the MIP promoter. Using DNase I footprinting analysis we found that purified Sp1 and AP2 transcription factors interact with several domains of the human MIP promoter sequence -253/+42. Furthermore, addition of purified Sp1 to Drosophila nuclear extracts activates in vitro transcription from the MIP promoter -253/+42. This promoter activity is competed by oligonucleotides containing domains footprinted with Sp1. Using promoter-reporter gene ( CAT ) constructs we found that the sequence -39/-70 contains a cis regulatory element essential for promoter activity in transient assays in lens cells. EMSA analysis showed that lens nuclear extracts contain factors that bind to the MIP 5'-flanking sequence containing overlapping Sp1 and AP2 ...Continue Reading

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Citations

May 4, 2001·Molecular Reproduction and Development·Z J TuD T Kiang
Oct 21, 2000·Development, Growth & Differentiation·H Ogino, K Yasuda
Dec 19, 2012·Human Genomics·Konstantinos Sousounis, Panagiotis A Tsonis
Oct 2, 2007·Progress in Retinal and Eye Research·Ales Cvekl, Melinda K Duncan
Apr 29, 2005·American Journal of Medical Genetics. Part B, Neuropsychiatric Genetics : the Official Publication of the International Society of Psychiatric Genetics·Karen SøebyThomas Werge
Jul 30, 2005·Biochimica Et Biophysica Acta·Fernando SegadeDonald W Bowden
Jan 27, 1999·Developmental Biology·J A West-MaysT Williams
Jun 12, 2012·Biochemical and Biophysical Research Communications·Periyasamy PalsamyToshimichi Shinohara
Jun 28, 2001·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·H YangS C Lu

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