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Volume 272, Number 27, Issue of July 4, 1997 pp. 16761-16768
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.

Rapid Protein Sequencing by Tandem Mass Spectrometry and cDNA Cloning of p20-CGGBP
A NOVEL PROTEIN THAT BINDS TO THE UNSTABLE TRIPLET REPEAT 5-d(CGG)_n-3 IN THE HUMAN FMR1 GENE

(Received for publication, February 19, 1997, and in revised form, April 24, 1997)

Heidrun Deissler , Matthias Wilm ^§ , Bülent Genç , Birgit Schmitz , Thomas Ternes , Frauke Naumann , Matthias Mann ^§ and Walter Doerfler

From the  Institut für Genetik, Universität zu Köln, D-50931 Köln and the ^§ Protein & Peptide Group, European Molecular Biology Laboratory, D-69117 Heidelberg, Federal Republic of Germany

The autonomous expansion of the unstable 5-d(CGG)_n-3 repeat in the 5-untranslated region of the human FMR1 gene leads to the fragile X syndrome, one of the most frequent causes of mental retardation in human males. We have recently described the isolation of a protein p20-CGGBP that binds sequence-specifically to the double-stranded trinucleotide repeat 5-d(CGG)-3 (Deissler, H., Behn-Krappa, A., and Doerfler, W. (1996) J. Biol. Chem. 271, 4327-4334). We demonstrate now that the p20-CGGBP can also bind to an interrupted repeat sequence. Peptide sequence tags of p20-CGGBP obtained by nanoelectrospray mass spectrometry were screened against an expressed sequence tag data base, retrieving a clone that contained the full-length coding sequence for p20-CGGBP. A bacterially expressed fusion protein p20-CGGBP-6xHis exhibits a binding pattern to the double-stranded 5-d(CGG)_n-3 repeat similar to that of the authentic p20-CGGBP. This novel protein lacks any overall homology to other known proteins but carries a putative nuclear localization signal. The p20-CGGBP gene is conserved among mammals but shows no homology to non-vertebrate species. The gene encoding the sequence for the new protein has been mapped to human chromosome 3.

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