title: The genomic region encompassing the nephropathic cystinosis gene (CTNS): complete sequencing of a 200-kb segment and discovery of a novel gene within the common cystinosis-causing deletion.
abstract: Nephropathic cystinosis is an autosomal recessive disorder caused by the defective transport of cystine out of lysosomes. Recently, the causative gene (CTNS) was identified and presumed to encode an integral membrane protein called cystinosin. Many of the disease-associated mutations in CTNS are deletions, including one >55 kb in size that represents the most common cystinosis allele encountered to date. In an effort to determine the precise genomic organization of CTNS and to gain sequence-based insight about the DNA within and flanking cystinosis-associated deletions, we mapped and sequenced the region of human chromosome 17p13 encompassing CTNS. Specifically, a bacterial artificial chromosome (BAC)-based physical map spanning CTNS was constructed by sequence-tagged site (STS)-content mapping. The resulting BAC contig provided the relative order of 43 STSs. Two overlapping BACs, which together contain all of the CTNS exons as well as extensive amounts of flanking DNA, were selected and subjected to shotgun sequencing. A total of 200,237 bp of contiguous, high-accuracy sequence was generated. Analysis of the resulting data revealed a number of interesting features about this genomic region, including the long-range organization of CTNS, insight about the breakpoints and intervening DNA associated with the common cystinosis-causing deletion, and structural information about five genes neighboring CTNS (human ortholog of rat vanilloid receptor subtype 1 gene, CARKL, TIP-1, P2X5, and HUMINAE). In particular, sequence analysis detected the presence of a novel gene (CARKL) residing within the most common cystinosis-causing deletion. This gene encodes a previously unknown protein that is predicted to function as a carbohydrate kinase. Interestingly, both CTNS and CARKL are absent in nearly half of all cystinosis patients (i.e., those homozygous for the common deletion). [The sequence data described in this paper have been submitted to the GenBank data library under accession nos. AF168787 and AF163573.]
authors.affiliation: NIH Intramural Sequencing Center, National Institutes of Health, Gaithersburg, Maryland 20877, USA.
authors.forename: J W
authors.initials: JW
authors.lastname: Touchman
authors.forename: Y
authors.initials: Y
authors.lastname: Anikster
authors.forename: N L
authors.initials: NL
authors.lastname: Dietrich
authors.forename: V V
authors.initials: VV
authors.lastname: Maduro
authors.forename: G
authors.initials: G
authors.lastname: McDowell
authors.forename: V
authors.initials: V
authors.lastname: Shotelersuk
authors.forename: G G
authors.initials: GG
authors.lastname: Bouffard
authors.forename: S M
authors.initials: SM
authors.lastname: Beckstrom-Sternberg
authors.forename: W A
authors.initials: WA
authors.lastname: Gahl
authors.forename: E D
authors.initials: ED
authors.lastname: Green
mesh_terms: D026921:Amino Acid Transport Systems, Neutral; D000818:Animals; D002478:Cells, Cultured; D002874:Chromosome Mapping; D002886:Chromosomes, Human, Pair 17 / Q000235:genetics; D003001:Cloning, Molecular; D003554:Cystinosis / Q000209:etiology / Q000235:genetics*; D006023:Glycoproteins; D006801:Humans; D019169:Jurkat Cells; D008565:Membrane Proteins / Q000235:genetics*; D026901:Membrane Transport Proteins; D008969:Molecular Sequence Data; D005810:Multigene Family; D010770:Phosphotransferases / Q000235:genetics*; D017853:Phosphotransferases (Alcohol Group Acceptor); D020161:Physical Chromosome Mapping; D051381:Rats; D017422:Sequence Analysis, DNA; D017384:Sequence Deletion / Q000235:genetics*; D014157:Transcription Factors / Q000235:genetics*; D014407:Tumor Cells, Cultured
chemical_list: D026921:Amino Acid Transport Systems, Neutral; C111445:CTNS protein, human; D006023:Glycoproteins; D008565:Membrane Proteins; D026901:Membrane Transport Proteins; D014157:Transcription Factors; D010770:Phosphotransferases; D017853:Phosphotransferases (Alcohol Group Acceptor); C404793:SHPK protein, human