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J. Biol. Chem., Vol. 279, Issue 24, 25653-25664, June 11, 2004

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Identification and Characterization of ZFP-57, a Novel Zinc Finger Transcription Factor in the Mammalian Peripheral Nervous System^*

María B. Durán Alonso, Georg Zoidl¶, Carla Taveggia||, Frank Bosse**, Christiane Zoidl¶, Mary Rahman, Eric Parmantier, Charlotte H. Dean, Brett S. Harris¶¶, Lawrence Wrabetz||, Hans Werner Müller**, Kristjan R. Jessen, and Rhona Mirsky||||

From the Department of Anatomy and Developmental Biology, University College London, Gower Street, London WC1E 6BT, United Kingdom, the ^¶Department of Neuroanatomy and Molecular Brain Research, Ruhr-University Bochum, University Street, 44780 Bochum, Germany, the ^||San Raffaele Scientific Institute, DIBIT, Via Olgettina 58, 20132 Milan, Italy, the ^**Molecular Neurobiology Laboratory, Department of Neurology, Heinrich-Heine-University, Moorenstrasse 5, D-40225 Düsseldorf, Germany, and the ^¶¶Department of Cell Biology and Anatomy, Medical University of South Carolina, Charleston, South Carolina 29425

To isolate new zinc finger genes expressed at early stages of peripheral nerve development, we have used PCR to amplify conserved zinc finger sequences. RNA from rat embryonic day 12 and 13 sciatic nerves, a stage when nerves contain Schwann cell precursors, was used to identify several genes not previously described in Schwann cells. One of them, zinc finger protein (ZFP)-57, proved to be the homologue of a mouse gene found in F9 teratocarcinoma cells. Its mRNA expression profile within embryonic and adult normal and transected peripheral nerves, and its distribution in the rest of the nervous system is described. High levels of expression are seen in embryonic nerves and spinal cord. These drop rapidly during the first few weeks after birth, a pattern mirrored in other parts of the nervous system. ZFP-57 localizes to the nucleus of Schwann and other cells. The sequence contains an N-terminal Krüppel-associated box (KRAB) domain and ZFP-57 constructs containing green fluorescent protein reveal that the protein colocalizes with heterochromatin protein 1 to centromeric heterochromatin in a characteristic speckled pattern in NIH3T3 cells. The KRAB domain is required for this localization, because constructs lacking it target the protein to the nucleus but not to the centromeric heterochromatin. When fused to a heterologous DNA binding domain, the KRAB domain of ZFP-57 represses transcription, and full-length ZFP-57 represses Schwann cell transcription from myelin basic protein and P₀ promoters in co-transfection assays. Zfp-57 mRNA is up-regulated in Schwann cells in response to leukemia inhibitory factor and fibroblast growth factor 2.

Received for publication, January 14, 2004 , and in revised form, April 5, 2004.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBank^TM/EBI Data Bank with accession number(s) AY344233.

^* This work was supported in part by a grant (to K. R. J. and R. M.) from the Wellcome Trust. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Both authors contributed equally to this work.

Current address: Oncology Disease Group, Aventis Pharma, 13 Quai Jules Guesde, 94403 Vitry-sur-Seine, France.

Current address: Howard Hughes Medical Institute and Developmental Biology Program, Memorial Sloan-Kettering Center, New York, NY 10021.

^|||| To whom correspondence should be addressed: Dept. of Anatomy and Developmental Biology, University College London, Gower Street, London WC1E 6BT, UK. Tel.: 44-20-7679-3380; Fax: 44-20-7679-2091; E-mail: r.mirsky{at}ucl.ac.uk.

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