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Initiation of zebrafish haematopoiesis by the TATA-box-binding protein-related factor Trf3

Nature volume 450, pages 1082–1085 (2007)Cite this article

Abstract

TATA-box-binding protein (TBP)-related factor 3, TRF3 (also called TBP2), is a vertebrate-specific member of the TBP family that has a conserved carboxy-terminal region and DNA-binding domain virtually identical to that of TBP (ref. 1). TRF3 is highly expressed during embryonic development, and studies in zebrafish and Xenopus have shown that it is required for normal embryogenesis2,3. Here we show that zebrafish embryos depleted of Trf3 exhibit multiple developmental defects and, in particular, fail to undergo haematopoiesis. Expression profiling for Trf3-dependent genes identified mespa, which encodes a transcription factor whose murine orthologue is required for mesoderm specification4, and chromatin immunoprecipitation verified that Trf3 binds to the mespa promoter. Depletion of Mespa resulted in developmental and haematopoietic defects markedly similar to those induced by Trf3 depletion. Injection of mespa messenger RNA (mRNA) restored normal development to a Trf3-depleted embryo, indicating mespa is the single Trf3 target gene required for zebrafish embryogenesis. Zebrafish embryos depleted of Trf3 or Mespa also failed to express cdx4, a caudal-related gene required for haematopoiesis. Mespa binds to the cdx4 promoter, and epistasis analysis revealed an ordered trf3–mespa–cdx4 pathway. Thus, in zebrafish, commitment of mesoderm to the haematopoietic lineage occurs through a transcription factor pathway initiated by a TBP-related factor.

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Figure 1: mespa is the single Trf3 target gene required for proper embryonic development.
Figure 2: Trf3- and Mespa-depleted embryos fail to undergo haematopoiesis.
Figure 3: Trf3 initiates a transcription factor pathway required for cdx4 expression and haematopoiesis.

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ArrayExpress

Data deposits

The microarray data have been deposited in the ArrayExpress database at http://www.ebi.ac.uk/arrayexpress under the accession number E-MEXP-1279.

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Acknowledgements

We thank C. Sagerstrom and M. Tsang for reagents, the Kimmel Cancer Center Microarray Core Facility at Thomas Jefferson University for performing the microarray analysis, members of the Lawson laboratory for technical support, and S. Evans for editorial assistance. The Zebrafish International Resource Center is supported by a grant from the National Institutes of Health – National Center for Research Resources (NIH-NCRR). This work was supported in part by a grant from the National Institutes of Health to M.R.G. M.R.G. is an investigator of the Howard Hughes Medical Institute.

Author Contributions D.O.H., N.D.L. and M.R.G. conceived and designed the experiments. D.O.H. performed the experiments, with the assistance of N.D.L., who performed the experiments shown in Supplementary Fig. 2, and T.R., who performed the chromatin immunoprecipitation assays shown in Fig. 1b and Supplementary Fig. 10a. D.O.H., T.R., N.D.L. and M.R.G. analysed the data. D.O.H., N.D.L. and M.R.G. wrote the paper.

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Authors and Affiliations

  1. Howard Hughes Medical Institute,

    Daniel O. Hart, Tamal Raha & Michael R. Green

  2. Programs in Gene Function and Expression and Molecular Medicine, University of Massachusetts Medical School, 364 Plantation Street, Worcester, Massachusetts 01605, USA ,

    Daniel O. Hart, Tamal Raha, Nathan D. Lawson & Michael R. Green

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  1. Daniel O. Hart
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Corresponding authors

Correspondence to Nathan D. Lawson or Michael R. Green.

Supplementary information

Supplementary Figures 1-10 and Supplementary Tables 1 and 2

This file contains 10 figures showing various control experiments that monitor trf3 and tbp expression during development, show Trf3- and Mespa-depleted embryos, confirm rescue of Trf3-depleted embryos following injection of mespa mRNA, and show Trf3 and Mespa are required for expression of several Cdx4-dependent hox genes. The file also contains 2 tables listing the genes that are significantly down-regulated upon Trf3 depletion and the sequences of all primers used in this paper. (PDF 1004 kb)

Supplementary Data

This file contains Microarray Data. The complete list of genes induced or repressed by trf3 MO treatment. (XLS 2330 kb)

Supplementary Corrigendum

The file contains Corrigendum for Supplementary Information. This file was uploaded on 13 December 2007 (PDF 21 kb)

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Hart, D., Raha, T., Lawson, N. et al. Initiation of zebrafish haematopoiesis by the TATA-box-binding protein-related factor Trf3. Nature 450, 1082–1085 (2007). https://doi.org/10.1038/nature06349

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