The cloning and characterization of a maternally expressed novel zinc finger nuclear phosphoprotein (xnf7) in Xenopus laevis.
We report the cloning of a cDNA (xnf7) coding for a maternally expressed Xenopus protein that becomes highly enriched in nuclei of the central nervous system during later development and in nuclei of adult brain. The protein also shows stage-specific nuclear/cytoplasmic partitioning and phosphorylation that may be related to its function. In addition, it binds to double-stranded DNA in vitro. The conceptual protein produced by the xnf7 clone contains several acidic domains, a novel zinc finger domain, three putative p34cdc2 protein kinase phosphorylation sites, and a bipartite basic nuclear localization signal. The xnf7 mRNA was detected as a maternal transcript that decreased in abundance during development through the gastrula stage. It was reexpressed at the neural stage in mesoderm and neural tissues, and its reexpression was not dependent upon the normal juxtaposition of the mesoderm and ectoderm that occurs during neural induction as demonstrated by high titer in exogastrulae. In situ hybridization showed enrichment of the mRNA in the neural tube and a small amount in the mesoderm at the late neurula stage. Xnf7 is normally phosphorylated during oocyte maturation. The bacterially expressed xnf7 protein was phosphorylated in vitro by purified maturation-promoting factor at a threonine in a small N-terminal domain containing one of the p34cdc2 protein kinase phosphorylation sites, but not by several other protein kinases. The structural domains present in the protein and its localization in nuclei suggest that the xnf7 gene product performs an important nuclear function during early development, perhaps as a transcription factor or a structural component of chromatin.
Department of Molecular Genetics, University of Texas M. D. Anderson Cancer Center, Houston 77030.
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Reddy
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Kloc
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Etkin
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D001921:Brain / Q000378:metabolism; D016203:CDC2 Protein Kinase / Q000378:metabolism; D003001:Cloning, Molecular; D046508:Culture Techniques; D004247:DNA; D004268:DNA-Binding Proteins; D004527:Egg Proteins; D004926:Escherichia coli / Q000235:genetics; D005260:Female; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000235:genetics*; D009693:Nucleic Acid Hybridization; D010750:Phosphoproteins / Q000235:genetics*; D010766:Phosphorylation; D014158:Transcription, Genetic; D029867:Xenopus Proteins; D014982:Xenopus laevis; D016335:Zinc Fingers / Q000235:genetics*
D004268:DNA-Binding Proteins; D004527:Egg Proteins; D009687:Nuclear Proteins; D010750:Phosphoproteins; D029867:Xenopus Proteins; C071551:XNF7 protein, Xenopus; D004247:DNA; D016203:CDC2 Protein Kinase