X-linked retinitis pigmentosa (XLRP) results from mutations in at least two different loci, designated RP2 and RP3, located at Xp11.3 and Xp21.1, respectively. The RP3 gene was recently isolated by positional cloning, whereas the RP2 locus was mapped genetically to a 5-cM interval. We have screened this region for genomic rearrangements by the YAC representation hybridization (YRH) technique and detected a LINE1 (L1) insertion in one XLRP patient. The L1 retrotransposition occurred in an intron of a novel gene that consisted of five exons and encoded a polypeptide of 350 amino acids. Subsequently, nonsense, missense and frameshift mutations, as well as two small deletions, were identified in six additional patients. The predicted gene product shows homology with human cofactor C, a protein involved in the ultimate step of ß-tubulin folding. Our data provide evidence that mutations in this gene, designated RP2, are responsible for progressive retinal degeneration.
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Bhattacharya, S.S. et al. Close genetic linkage between X-linked retinitis pigmentosa and a restriction fragment length polymorphism identified by recombinant DNA probe L1.28. Nature 309, 253–255 ( 1984).
Wirth, B. et al. Two different genes for X-linked retinitis pigmentosa. Genomics 2, 263–266 ( 1988).
Ott, J. et al. Localizing multiple X chromosome linked retinitis pigmentosa loci using multilocus homogeneity tests. Proc. Natl Acad. Sci. USA 87, 701 –704 (1990).
Teague, P.W. Heterogeneity analysis in 40 X-linked retinitis pigmentosa families. Am. J. Hum. Genet. 55, 105–111 ( 1994).
Meindl, A. et al. A gene (RPGR) with homology to the RCC1 guanine nucleotide exchange factor is mutated in X-linked retinitis pigmentosa (RP3). Nature Genet. 13, 36–42 ( 1996).
Roepman, R. et al. Positional cloning of the gene for X-linked retinitis pigmentosa 3: homology with the guanine-nucleotide-exchange factor RCC1. Hum. Mol. Genet. 5, 1035–1041 ( 1996).
Buraczynska, M. et al. Spectrum of mutations in the RPGR gene that are identified in 20% of families with X-linked retinitis pigmentosa. Am. J. Hum. Genet. 61, 1287–1292 ( 1997).
Thiselton, D.L. et al. Mapping the RP2 locus for X-linked retinitis pigmentosa on proximal Xp: a genetically defined 5-cM critical region and exclusion of candidate genes by physical mapping. Genome Res. 6, 1093–1102 (1996).
Roepman, R. et al. Identification of a gene disrupted by a microdeletion in a patient with X-linked retinitis pigmentosa (XLRP). Hum. Molec. Genet. 5, 827–833 (1996).
Tian, G. et al. Pathway leading to correctly folded ß-tubulin. Cell 86, 287–296 (1996).
Kazazian, H.H. & Moran, J.V.J. The impact of L1 retrotransposons on the human genome. Nature Genet. 19, 19 –24 (1998).
Economou-Pachnis, A., Lohse, M.A., Furano, A.V. & Tsichlis, P.N. Insertion of long interspersed repeated elements at the Igh (immunoglobulin heavy chain) and Mlvi-2 (Moloney leukemia virus integration 2) loci of rat . Proc. Natl Acad. Sci. USA 9, 2857– 2861 (1985).
Boccaccio, C., Deschatrette, J. & Meunier-Rotival, M. Empty and occupied insertion site of the truncated LINE-1 repeat located in the mouse serum albumin-encoding gene. Gene 88, 181–186 (1990).
Frantz, S.A., Thiara, A.S., Lodwick, D. & Samani, N.J. A major polymorphism in the rat SA gene caused by the insertion of a LINE element. Mamm. Genome 7, 865–866 (1996).
Kazazian, H.H. et al. Haemophilia A resulting from de novo insertion of L1 sequences represents a novel mechanism for mutation in man. Nature 332, 164–166 (1988).
Miki, Y. et al. Disruption of the APC gene by a retrotransposal insertion of L1 sequence in a colon cancer. Cancer Res. 52, 643– 645 (1992).
Narita, N. et al. Insertion of a 5´ truncated L1 element into the 3´ end of exon 44 of the dystrophin gene resulted in skipping of the exon during splicing in a case of Duchenne muscular dystrophy. J. Clin. Invest. 91, 1862–1867 (1993).
Mulhardt, C. et al. The spastic mouse: aberrant splicing of glycine receptor beta subunit mRNA caused by intronic insertion of L1 element. Neuron 13, 1003–1015 (1994).
Takahara, T. et al. Dysfunction of the Orleans reeler gene arising from exon skipping due to transposition of a full-length copy of an active L1 sequence into the skipped exon. Hum. Molec. Genet. 5, 989–993 (1996).
Perou, C.M. et al. Identification of the murine beige gene by YAC complementation and positional cloning. Nature Genet. 13, 303 –308 (1996).
Cremers, F.P.M., van de Pol, D.J.R., van Kerkhoff, L.P.M., Wieringa, B. & Ropers, H.H. Cloning of a gene that is rearranged in patients with choroideremia. Nature 347, 674–677 (1990).
McCarthy, J.E.G. & Kollmus, H. Cytoplasmic mRNA-protein interactions in eukaryotic gene expression. Trends Biochem. Sci. 20, 191–197 (1995).
Ross, J. Control of messenger RNA stability in higher eukaryotes. Trends Genet. 12 , 171–175 (1996).
Woodford, B.J. & Blanks, J.C. Localization of actin and tubulin in dveloping and adult mammalian photoreceptors. Cell & Tissue Res. 256, 495–505 ( 1989).
Watanabe, M., Rutishauser, U. & Silver, J. Formation of the retinal ganglion cell and optic fiber layers. J. Neurobiol. 22, 85–96 (1991).
Sale, W.S., Besharse, J.C. & Piperno, G. Distribution of acetylated alpha-tubulin in retina and in vitro-assembled microtubules. Cell Motil. Cytoskeleton 9, 243–253 (1988).
Lewis, G.P., Matsumoto, B. & Fisher, S.K. Changes in the organization and expression of cytoskeletal proteins during retinal degeneration induced by retinal detachment. Invest. Ophthalmol. Vis. Sci. 36, 2404–2416 (1995).
Muresan, V., Joshi, H.C. & Besharse, J.C. Gamma-tubulin in differentiated cell types: localization in the vicinity of basal bodies in retinal photoreceptors and ciliated epithelia . J. Cell Sci. 104, 1229– 1237 (1993).
Coleman, M.P. et al. 1.8-Mb YAC contig in Xp11.23: identification of CpG islands and physical mapping of CA repeats in a region of high gene density. Genomics 21, 337–343 (1994).
Hagemann, T. et al. Physical mapping in a YAC contig of 11 markers on the human X chromosome in Xp11.23. Genomics 21, 262– 265 (1994).
Mueller, P.R. & Wold, B. In vivo footprinting of a muscle specific enhancer by ligation mediated PCR. Science 246, 780–786 (1989).
Skowronski, J. & Singer, M.F. The abundant LINE-1 family of repeated DNA sequences in mammals: genes and pseudogenes. Cold Spring Harb. Symp. Quant. Biol. LI, 457–464 (1986).
Siebert, P.D., Chenchik, A., Kellogg, D.E., Lukyanov, K.A. & Lukyanov, S.A. An improved PCR method for walking in uncloned genomic DNA. Nucleic Acids Res. 23, 1087– 1088 (1995).
Berger, W. et al. Isolation of a candidate gene for Norrie disease by positional cloning. Nature Genet. 1, 199–203 (1992).
Ishihara, A., Gee, K., Schwartz, S., Jacobson, K. & Lee, J. Direct hybridization of large-insert genomic clones on high-density gridded cDNA filter arrays. Biotechniques 23, 120–124 (1997).
Adam, G., Miller, S., Flam, F. & Ohlsson, R. Allele-specific in situ hybridization (ASISH) analysis: a novel technique which resolves differential allelic usage of H19 within the same cell lineage during human placental development. Development 122, 839–847 (1996).
The authors are very grateful to the patients and their families who contributed to this study. We would like to thank the Ressourcenzentrum im Deutschen Humangenomprojekt, the UK HGMP Resource Center (Hinxton) and the YAC Screening Center Leiden, The Netherlands, for providing us with genomic and cDNA libraries. We also wish to thank S. Freier and S.v.d. Velde-Visser for tissue culturing, C. Zeitz and M. Krause for technical assistance, M.R. Toliat for assistance during characterization of the cDNA and A.d. Hollander for making available total RNA from adult human retina. This work was supported in part by the Deutsche Forschungsgemeinschaft (SL, grant Be 1559/2-1) and the Foundation Fighting Blindness, USA (R.K.).
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Schwahn, U., Lenzner, S., Dong, J. et al. Positional cloning of the gene for X-linked retinitis pigmentosa 2. Nat Genet 19, 327–332 (1998). https://doi.org/10.1038/1214
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