REPRESSOR ELEMENTS IN THE CODING REGION OF THE HUMAN HISTONE H4 GENE INTERACT WITH THE TRANSCRIPTION FACTOR CDP CUT/

The coding region of the human histone H4 gene FO108 undergoes dynamic changes in chromatin structure that correlate with modifications in g ene expression. Such structural alterations generally reflect transcri ption factor interactions with gene regulatory sequences. To test for regulatory elements within the coding region, we performed transient t ransfection experiments in HeLa cells using constructs with histone H4 sequences fused upstream of a heterologous thymidine kinase promoter and CAT reporter gene. H4 gene sequences from -10 to +210 repressed tr anscription 4.8-fold. Further deletion and mutational analysis delinea ted three repressor elements within this region. Using oligonucleotide competition analysis and specific antibody recognition in electrophor etic mobility shift assays, as well as methylation interference and DN ase I footprinting analyses, we have identified the CCAAT displacement protein (CDP/cut) as the factor that interacts with these three repre ssor elements. CDP/cut binding to these repressor sites is proliferati on-specific and cell-cycle-regulated, increasing in mid to late S phas e. Our results indicate that the proximal 200 nucleotides of the histo ne H4-coding region contain transcriptional regulatory elements that m ay contribute to cell-cycle control of histone gene expression by inte racting with repressor complexes containing CDP/cut homeodomain transc ription factors. (C) 1998 Published by Elsevier Science B.V. All right s reserved.