Nucleic Acids Research, 1995, Vol. 23, No. 13 2389-2395
© 1995
MOLECULAR BIOLOGY |
Cloning and characterisation of a nuclear, site specific ssDNA binding protein
Laboratory of Biochemistry, university of Groningen Nijenbourgh 4, 9747 AG Groningen, The Netherlands
*To whom corespondence should be addressed
Received April 19, 1995. Accepted May 24, 1995.
Estradlol Inducibte, liver-specific expression of the apoVLDL II gene is mediated through the estrogen receptor and a variety of other DNA-binding proteins. In the present study we report the cloning and characterisation of a single-strand DNA binding protein that interacts with the lower strand of a complex regulatory site, which includes the major estrogen responsive element and a site that resembles the rat albumin site D (apoVLDL II site D). Based on its binding specificity etermined with electro-mobility shift assays, the protein is named single-strand D-box binding factor (ssDBF). Analysis of the deduced 302 amino acid sequence revealed that the protein belongs to the eteronuclear ribonucleoprotein A/B family (hnRNP A/B) and resembles other known eukaryotic single-strand DNA binding proteins. Transient transfecf on experiments in a chicken liver cell-line showed that the protein represses estrogen-Induced transcription. A protein with similar binding characteristics is present in liver nuclear extract. The relevance of the occurrence of this protein to the expression of the apoVLDL II gene Is discussed.
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