Transcriptional element H4-site II of cell cycle regulated human H4 histone genes is a multipartite protein/DNA interaction site for factors HiNF-D, HiNF-M, and HiNF-P: involvement of phosphorylation
Graduate School of Biomedical Sciences; Department of Cell Biology; Department of Medicine, Diabetes Division
Medical Subject Headings
Base Sequence; Binding Sites; Cell Cycle; Chromatography, Affinity; DNA-Binding Proteins; Histones; Humans; Molecular Sequence Data; Phosphoprotein Phosphatases; Phosphorylation; Plasmids; Protein Processing, Post-Translational; Transcription Factors; *Transcription, Genetic
Life Sciences | Medicine and Health Sciences
Cell cycle regulated gene expression was studied by analyzing protein/DNA interactions occurring at the H4-Site II transcriptional element of H4 histone genes using several approaches. We show that this key proximal promoter element interacts with at least three distinct sequence-specific DNA binding activities, designated HiNF-D, HiNF-M, and HiNF-P. HiNF-D binds to an extended series of nucleotides, whereas HiNF-M and HiNF-P recognize sequences internal to the HiNF-D binding domain. Gel retardation assays show that HiNF-D and HiNF-M each are represented by two distinct protein/DNA complexes involving the same DNA binding activity. These results suggest that these factors are subject to post-translational modifications. Dephosphorylation experiments in vitro suggest that both electrophoretic mobility and DNA binding activities of HiNF-D and HiNF-M are sensitive to phosphatase activity. We deduce that these factors may require a basal level of phosphorylation for sequence specific binding to H4-Site II and may represent phosphoproteins occurring in putative hyper- and hypo-phosphorylated forms. Based on dramatic fluctuations in the ratio of the two distinct HiNF-D species both during hepatic development and the cell cycle in normal diploid cells, we postulate that this modification of HiNF-D is related to the cell cycle. However, in several tumor-derived and transformed cell types the putative hyperphosphorylated form of HiNF-D is constitutively present. These data suggest that deregulation of a phosphatase-sensitive post-translational modification required for HiNF-D binding is a molecular event that reflects abrogation of a mechanism controlling cell proliferation. Thus, phosphorylation and dephosphorylation of histone promoter factors may provide a basis for modulation of protein/DNA interactions and H4 histone gene transcription during the cell cycle and at the onset of quiescence and differentiation.
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Citation: J Cell Biochem. 1991 Jun;46(2):174-89. Link to article on publisher's site
Journal of cellular biochemistry
Van Wijnen, Andre J.; Ramsey-Ewing, Anna L.; Bortell, Rita; Owen, T. A.; Lian, Jane B.; Stein, Janet L.; and Stein, Gary S., "Transcriptional element H4-site II of cell cycle regulated human H4 histone genes is a multipartite protein/DNA interaction site for factors HiNF-D, HiNF-M, and HiNF-P: involvement of phosphorylation" (1991). GSBS Student Publications. 1291.