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

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),-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.