Nucleic Acids Research, Vol 25, Issue 7 1333-1338, Copyright © 1997 by Oxford University Press
A Bevilacqua, MT Fiorenza and F Mangia
To investigate the control of zygotic genome expression in two-cell mouse
embryos, we studied transcription factors required for transient expression
of microinjected DNA constructs driven by the promoter of one of the
earliest genes activated after fertilization in this system, the heat shock
gene hsp70. Cis-acting elements required for hsp70 activation were first
investigated by mutational analysis. Mutation of the TATA box and a
proximal GC box strongly inhibited construct expression, while that of a
CCAAT box had no effect. Transcription factors binding the wild-type hsp70
promoter were then titrated in vivo by coinjecting the construct with
double-stranded oligodeoxyribonucleotides containing definite consensus
sequences. Wild- type GC box oligonucleotides strongly inhibited construct
expression, while those containing mutated GC boxes, wild-type CCAAT boxes,
and heat shock elements had no effects. Finally, construct expression was
challenged by coinjecting antibodies to specific transcription factors.
Antibodies to factor Sp1 depressed construct expression in a dose-
dependent manner, while those to Sp2, HSF1 and HSF2 were ineffective. These
results pinpoint the Sp1 transcription factor as an absolute requirement
for activation of the hsp70 gene promoter in two-cell mouse embryos, and
make this factor a candidate for a major regulator of the onset of murine
zygotic genome expression.
ARTICLES
Developmental activation of an episomic hsp70 gene promoter in two-cell mouse embryos by transcription factor Sp1
Department of Psychology and Department of Histology and Medical Embryology, La Sapienza University of Rome, Via Borelli 50, 00161 Rome, Italy.
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