Interactions with Single-stranded and Double-stranded DNA-binding Factors and Alternative Promoter Conformation upon Transcriptional Activation of the Htf9-a/RanBP1 and Htf9-c Genes

doi:10.1074/jbc.273.1.495

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Vol. 273, Issue 1, 495-505, January 2, 1998

Interactions with Single-stranded and Double-stranded DNA-binding Factors and Alternative Promoter Conformation upon Transcriptional Activation of the Htf9-a/RanBP1 and Htf9-c Genes

Gigliola Di Matteo, Massimiliano Salerno, Giulia Guarguaglini, Barbara Di Fiore, Franco Palitti^**, and Patrizia Lavia

From the CNR Centre of Evolutionary Genetics, c/o Department of Genetics and Molecular Biology and the ^** Department of Biochemical Sciences, University "La Sapienza," Rome 00185, Italy

The murine Htf9-a/RanBP1 and Htf9-c genes are divergently transcribed from a shared TATA-less promoter. Transcription of bothgenes is initiated on complementary DNA strands and is controlledby cell cycle-dependent mechanisms. The bidirectional promoterharbors a genomic footprint flanking the major transcription startsite of both genes. Transient promoter assays showed that thefootprinted element is important for transcription of both genes.Protein-binding experiments and antibody assays indicated thatmembers of the retinoid X receptor family interact with the double-strandedsite. In addition, distinct factors interact with single DNA strandsof the element. Double-stranded binding factors were highly expressedin liver cells, in which neither gene is transcribed, while single-strandedbinding proteins were abundant in cycling cells, in which transcriptionof both genes is efficient. In vivo S1 analysis of the promoterdepicted an S1-sensitive organization in cells in which transcriptionof both genes is active; S1 sensitivity was not detected in conditionsof transcriptional repression. Thus, the same element is a targetfor either retinoid X receptor factors, or for single-strandedbinding proteins, and form distinct complexes in different cellularconditions depending on the DNA conformation in the binding site.

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