Human replication-dependent histone H3 genes are activated by a tandemly arranged pair of two CCAAT boxes.
We have analysed the transcriptional regulation of the human histone H3 genes using promoter deletion series, scanning mutagenesis, specific mutagenesis and electrophoretic mobility-shift assay experiments. The promoters of five of the six examined histone H3 genes showed near-maximal activity at lengths of 133-227 bp: H3/d 198 bp, H3/h 147 bp, H3/k 133 bp, H3/m 227 bp, H3/n 140 bp (exception H3/i). To search for functional cis-elements within these regions, we performed scanning mutagenesis of the two histone H3 promoters H3/k and H3/m. Mutagenesis revealed that the functional framework of the histone H3 promoters consists of a TATA box and two tandemly arranged CCAAT boxes in relatively fixed positions. Alterations of the distance between the CCAAT boxes and of the distance between the CCAAT boxes and the TATA box resulted in significant loss of activity. In electrophoretic mobility-shift assay experiments, the factor CBF (CCAAT-binding factor)/NF-Y (nuclear factor-Y) bound to isolated CCAAT boxes of the H3/k promoter. This suggests that an initiation complex is formed on the histone H3 promoter that has a defined structure and limited flexibility, consisting of two molecules of CBF/NF-Y and further (general or specific) transcription factors.
Institut für Biochemie und Molekulare Zellbiologie, Universität Göttingen, Humboldtallee 23, D-37073 Göttingen, Germany.
Heiner
H
Koessler
Joerg
J
Kahle
Christa
C
Bode
Detlef
D
Doenecke
Werner
W
Albig
D001483:Base Sequence / Q000235:genetics; D001665:Binding Sites / Q000235:genetics; D022762:CCAAT-Enhancer-Binding Proteins / Q000235:genetics*; D003001:Cloning, Molecular; D004247:DNA / Q000737:chemistry / Q000235:genetics; D004261:DNA Replication / Q000502:physiology*; D024202:Electrophoretic Mobility Shift Assay / Q000379:methods; D004742:Enhancer Elements, Genetic / Q000235:genetics; D005786:Gene Expression Regulation / Q000502:physiology*; D006657:Histones / Q000235:genetics* / Q000502:physiology; D006801:Humans; D007668:Kidney / Q000737:chemistry / Q000166:cytology / Q000196:embryology / Q000378:metabolism; D008969:Molecular Sequence Data; D016296:Mutagenesis / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics; D017422:Sequence Analysis, DNA / Q000379:methods; D020080:Tandem Repeat Sequences / Q000502:physiology*; D014158:Transcription, Genetic / Q000502:physiology
D022762:CCAAT-Enhancer-Binding Proteins; D006657:Histones; D004247:DNA