An octamer element is required for the expression of the alpha H2B histone gene during the early development of the sea urchin.
Early (alpha) histone genes are one of several histone gene families in the sea urchin genome. They are expressed at high levels in blastula-stage embryos and are inactivated by the early gastrula stage. By microinjecting mutant early H2B genes into sea urchin zygotes and monitoring their transcriptional activity in blastula- and gastrula-stage embryos, we sought to identify the cis-regulatory elements responsible for this dramatic change in early H2B gene activity. We found that deletion of DNA 5' of -71 and 3' of +591 did not affect the timing or magnitude of early H2B gene expression. Neither was early H2B gene expression affected by the replacement of sequences downstream of -36 with the corresponding region of the L1 late H2B gene, expressed after the peak transcription of the early H2B gene. Further deletion of early H2B promoter sequences from -71 to -56, removing a conserved octamer element, resulted in near-complete inactivation of the early H2B gene in both blastula- and gastrula-stage embryos. Also inactivating early H2B gene expression were an internal deletion of the octamer element and a base substitution mutation that altered its sequence. This base substitution mutation also caused a parallel reduction in the ability of the octamer element to bind a factor present in nuclear extracts of sea urchin blastulae. These data strongly suggest that the proper expression of the early H2B gene in cleavage- and blastula-stage embryos depends on the octamer element and a factor with which it interacts.
Bell
J
J
Department of Biochemistry, University of Southern California, School of Medicine, Los Angeles 90033.
Char
B R
BR
Maxson
R
R
D000818:Animals; D001483:Base Sequence; D002872:Chromosome Deletion; D003001:Cloning, Molecular; D005775:Gastrula / Q000502:physiology; D005786:Gene Expression Regulation; D006657:Histones / Q000235:genetics*; D008969:Molecular Sequence Data; D005810:Multigene Family; D016297:Mutagenesis, Site-Directed; D009838:Oligodeoxyribonucleotides; D015183:Restriction Mapping; D012617:Sea Urchins / Q000196:embryology* / Q000235:genetics; D012689:Sequence Homology, Nucleic Acid
D006657:Histones; D009838:Oligodeoxyribonucleotides