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Purification of the human histone H4 gene-specific transcription factors H4TF-1 and H4TF-2

Journal Article · · Genes Dev. (Cold Spring Harbor) (NY); (United States)
; ;
Transcription of the cell-cycle-regulated human histone genes increases approximately fivefold during S phase. One step toward the elucidation of the biochemical mechanisms that govern cell-cycle-regulated expression of these genes is to purify and characterize the transcription factors that regulate these promoters. Here, we describe the purification of two previously identified factors, H4TF-1 and H4TF-2, which bind the human histone H4 promoter. Purification was achieved through a combination of ion-exchange and oligonucleotide affinity chromatography. On the basis of analysis of purified fractions by SDS-polyacrylamide gels and UV cross-linking, we believe that H4TF-1 is two polypeptides of 105 and 110 kD. This factor binds to a GC-rich DNA sequence required for maximal expression of the H4 gene but does not bind to any Sp1 consensus elements tested. H4TF-2 is a 65-kD protein that binds specifically to sequences within that highly conserved H4 subtype-specific consensus promoter element. Both highly purified factors activated transcription in vitro only from H4 promoters that contained their binding sequences, demonstrating that H4TF-1 and H4TF-2 are H4-specific transcription factors that potentiate expression of this gene.
Research Organization:
Rockefeller Univ., New York, NY (USA)
OSTI ID:
5987131
Journal Information:
Genes Dev. (Cold Spring Harbor) (NY); (United States), Journal Name: Genes Dev. (Cold Spring Harbor) (NY); (United States) Vol. 2:12B; ISSN GEDEE
Country of Publication:
United States
Language:
English

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Related Subjects

560120* -- Radiation Effects on Biochemicals
Cells
& Tissue Culture
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
CHEMICAL RADIATION EFFECTS
CHEMICAL REACTIONS
CHEMISTRY
CHROMATOGRAPHY
CROSS-LINKING
DNA SEQUENCING
ELECTROMAGNETIC RADIATION
ELECTROPHORESIS
FRACTIONATION
GENE REGULATION
HELA CELLS
HISTONES
ION EXCHANGE CHROMATOGRAPHY
METHYLATION
MUTATIONS
NUCLEIC ACIDS
NUCLEOPROTEINS
OLIGONUCLEOTIDES
ORGANIC COMPOUNDS
PEPTIDES
POLYMERIZATION
PROTEINS
RADIATION CHEMISTRY
RADIATION EFFECTS
RADIATIONS
SEPARATION PROCESSES
STRUCTURAL CHEMICAL ANALYSIS
TRANSCRIPTION
ULTRAVIOLET RADIATION