Translation of Maternal TATA-Binding Protein mRNA Potentiates Basal but Not Activated Transcription in Xenopus Embryos at the Midblastula Transition
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Published in: | Molecular and Cellular Biology Vol. 19; no. 12; pp. 7972 - 7982 |
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AbstractList | 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. 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. Article Usage Stats Services MCB Citing Articles Google Scholar PubMed Related Content Social Bookmarking CiteULike Delicious Digg Facebook Google+ Mendeley Reddit StumbleUpon Twitter current issue Spotlights in the Current Issue MCB About MCB Subscribers Authors Reviewers Advertisers Inquiries from the Press Permissions & Commercial Reprints ASM Journals Public Access Policy MCB RSS Feeds 1752 N Street N.W. • Washington DC 20036 202.737.3600 • 202.942.9355 fax • journals@asmusa.org Print ISSN: 0270-7306 Online ISSN: 1098-5549 Copyright © 2014 by the American Society for Microbiology. For an alternate route to MCB .asm.org, visit: MCB |
Author | Gert Jan C. Veenstra Olivier H. J. Destrée Alan P. Wolffe |
AuthorAffiliation | 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 Netherlands 2 |
AuthorAffiliation_xml | – name: 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 Netherlands 2 |
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BackLink | https://www.ncbi.nlm.nih.gov/pubmed/10567523$$D View this record in MEDLINE/PubMed |
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Notes | ObjectType-Article-2 SourceType-Scholarly Journals-1 ObjectType-Feature-1 content type line 23 ObjectType-Article-1 ObjectType-Feature-2 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@helix.nih.gov. |
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Mendeley... Early embryonic development in Xenopus laevis is characterized by transcriptional repression which is relieved at the midblastula stage (MBT). Here we show... Early embryonic development in Xenopus laevis is characterized by transcriptional repression which is relieved at the midblastula stage (MBT). Here we show... |
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SubjectTerms | Animals Blastocyst - metabolism DNA-Binding Proteins - genetics DNA-Binding Proteins - metabolism Freshwater Gene Expression Regulation, Developmental Oct-1 protein Ovum Protein Biosynthesis RNA, Messenger - metabolism TATA-Box Binding Protein Transcription Factors - genetics Transcription Factors - metabolism Transcriptional Activation Transcriptional Regulation Xenopus laevis Xenopus laevis - embryology |
Title | Translation of Maternal TATA-Binding Protein mRNA Potentiates Basal but Not Activated Transcription in Xenopus Embryos at the Midblastula Transition |
URI | http://mcb.asm.org/content/19/12/7972.abstract https://www.tandfonline.com/doi/abs/10.1128/MCB.19.12.7972 https://www.ncbi.nlm.nih.gov/pubmed/10567523 https://search.proquest.com/docview/17431908 https://search.proquest.com/docview/69292195 https://pubmed.ncbi.nlm.nih.gov/PMC84882 |
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