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Molecular and Cellular Biology, December 1999, p. 7972-7982, Vol. 19, No. 12
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Translation of Maternal TATA-Binding Protein mRNA Potentiates
Basal but Not Activated Transcription in Xenopus Embryos
at the Midblastula Transition
Gert Jan C.
Veenstra,1
Olivier H. J.
Destrée,2 and
Alan P.
Wolffe1,*
Laboratory for Molecular Embryology, National
Institute of Child Health and Human Development, Bethesda, Maryland
20892,1 and Hubrecht Laboratory,
Netherlands Institute for Developmental Biology, 3584 CT Utrecht,
The Netherlands2
Received 11 June 1999/Returned for modification 3 August
1999/Accepted 7 September 1999
Early embryonic development in Xenopus laevis is
characterized by transcriptional repression which is relieved at the
midblastula stage (MBT). Here we show that the relative abundance of
TATA-binding protein (TBP) increases robustly at the MBT and that the
mechanism underlying this increase is translation of maternally
stored TBP RNA. We show that TBP is rate-limiting in egg extract under
conditions that titrate nucleosome assembly. Precocious
translation of TBP mRNA in Xenopus embryos facilitates
transcription before the MBT, without requiring TBP to be prebound to
the promoter before injection. This effect is transient in the absence
of chromatin titration and is sustained when chromatin is titrated.
These data show that translational regulation of TBP RNA contributes to
limitations on the transcriptional capacity before the MBT. Second, we
examined the ability of trans-acting factors to contribute
to promoter activity before the MBT. Deletion of cis-acting
elements does not affect histone H2B transcription in egg extract, a
finding indicative of limited trans-activation. Moreover,
in the context of the intact promoter, neither the transcriptional
activator Oct-1, nor TBP, nor TFIID enable transcriptional activation
in vitro. HeLa cell extract, however, reconstitutes activated
transcription in mixed extracts. These data suggest a deficiency in egg
extract cofactors required for activated transcription. We show
that the capacity for activated H2B transcription is gradually acquired at the early gastrula transition. This transition occurs well after the
blastula stage when the basal transcription machinery can first be
complemented with TBP.
*
Corresponding author. Mailing address: Laboratory for
Molecular Embryology, NICHHD, NIH, Bldg. 18T, Rm. 106, Bethesda, MD 20892. Phone: (301) 496-4045. Fax: (301) 402-1323. E-mail:
awlme{at}helix.nih.gov.
Molecular and Cellular Biology, December 1999, p. 7972-7982, Vol. 19, No. 12
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
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