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Molecular and Cellular Biology, September 1998, p. 5557-5566, Vol. 18, No. 9
0270-7306/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

The Maternal CCAAT Box Transcription Factor Which Controls GATA-2 Expression Is Novel and Developmentally Regulated and Contains a Double-Stranded-RNA-Binding Subunit

Robert L. Orford,1 Carl Robinson,1 Joanna M. Haydon,1 Roger K. Patient,2 and Matthew J. Guille1,*

Biophysics Laboratories, Division of Molecular and Cell Biology, Institute of Biomolecular and Biomedical Sciences, University of Portsmouth, Portsmouth PO1 2DY,1 and Developmental Biology Research Centre, Biomedical Sciences Division, The Randall Institute, King's College London, London WC2B 5RL,2 United Kingdom

Received 17 February 1998/Returned for modification 24 March 1998/Accepted 10 June 1998

The transcription factor GATA-2 is expressed at high levels in the nonneural ectoderm of the Xenopus embryo at neurula stages, with lower amounts of RNA present in the ventral mesoderm and endoderm. The promoter of the GATA-2 gene contains an inverted CCAAT box conserved among Xenopus laevis, humans, chickens, and mice. We have shown that this sequence is essential for GATA-2 transcription during early development and that the factor binding it is maternal. The DNA-binding activity of this factor is detectable in nuclei and chromatin bound only when zygotic GATA-2 transcription starts. Here we report the characterization of this factor, which we call CBTF (CCAAT box transcription factor). CBTF activity mainly appears late in oogenesis, when it is nuclear, and the complex has multiple subunits. We have identified one subunit of the factor as p122, a Xenopus double-stranded-RNA-binding protein. The p122 protein is perinuclear during early embryonic development but moves from the cytoplasm into the nuclei of embryonic cells at stage 9, prior to the detection of CBTF activity in the nucleus. Thus, the accumulation of CBTF activity in the nucleus is a multistep process. We show that the p122 protein is expressed mainly in the ectoderm. Expression of p122 mRNA is more restricted, mainly to the anterior ectoderm and mesoderm and to the neural tube. Two properties of CBTF, its dual role and its cytoplasm-to-nucleus translocation, are shared with other vertebrate maternal transcription factors and may be general properties of these proteins.

* Corresponding author. Mailing address: Division of Molecular and Cell Biology, School of Biological Sciences, University of Portsmouth, King Henry Building, King Henry I St., Portsmouth PO1 2DY, U.K. Phone: 01705 842066. Fax: 01705 842053. E-mail: matthew.guille{at}port.ac.uk.

Molecular and Cellular Biology, September 1998, p. 5557-5566, Vol. 18, No. 9
0270-7306/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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