title: Functional analysis of mutations in SLC7A9, and genotype-phenotype correlation in non-Type I cystinuria.
abstract: Cystinuria (OMIM 220100) is a common recessive disorder of renal reabsorption of cystine and dibasic amino acids that results in nephrolithiasis of cystine. Mutations in SLC3A1, which encodes rBAT, cause Type I cystinuria, and mutations in SLC7A9, which encodes a putative subunit of rBAT (b(o,+)AT), cause non-Type I cystinuria. Here we describe the genomic structure of SLC7A9 (13 exons) and 28 new mutations in this gene that, together with the seven previously reported, explain 79% of the alleles in 61 non-Type I cystinuria patients. These data demonstrate that SLC7A9 is the main non-Type I cystinuria gene. Mutations G105R, V170M, A182T and R333W are the most frequent SLC7A9 missense mutations found. Among heterozygotes carrying these mutations, A182T heterozygotes showed the lowest urinary excretion values of cystine and dibasic amino acids. Functional analysis of mutation A182T after co-expression with rBAT in HeLa cells revealed significant residual transport activity. In contrast, mutations G105R, V170M and R333W are associated to a complete or almost complete loss of transport activity, leading to a more severe urinary phenotype in heterozygotes. SLC7A9 mutations located in the putative transmembrane domains of b(o,+)AT and affecting conserved amino acid residues with a small side chain generate a severe phenotype, while mutations in non-conserved residues give rise to a mild phenotype. These data provide the first genotype-phenotype correlation in non-Type I cystinuria, and show that a mild urinary phenotype in heterozygotes may associate with mutations with significant residual transport activity.
authors.affiliation: Centre de Genètica Mèdica i Molecular (IRO), Hospital Duran i Reynals, Autovía de Castelldefels Km 2.7, L'Hospitalet de Llobregat, E-08907 Barcelona, Spain.
authors.forename: M A
authors.initials: MA
authors.lastname: Font
authors.forename: L
authors.initials: L
authors.lastname: Feliubadaló
authors.forename: X
authors.initials: X
authors.lastname: Estivill
authors.forename: V
authors.initials: V
authors.lastname: Nunes
authors.forename: E
authors.initials: E
authors.lastname: Golomb
authors.forename: Y
authors.initials: Y
authors.lastname: Kreiss
authors.forename: E
authors.initials: E
authors.lastname: Pras
authors.forename: L
authors.initials: L
authors.lastname: Bisceglia
authors.forename: A P
authors.initials: AP
authors.lastname: d'Adamo
authors.forename: L
authors.initials: L
authors.lastname: Zelante
authors.forename: P
authors.initials: P
authors.lastname: Gasparini
authors.forename: M T
authors.initials: MT
authors.lastname: Bassi
authors.forename: A L
authors.initials: AL
authors.lastname: George
authors.forename: M
authors.initials: M
authors.lastname: Manzoni
authors.forename: M
authors.initials: M
authors.lastname: Riboni
authors.forename: A
authors.initials: A
authors.lastname: Ballabio
authors.forename: G
authors.initials: G
authors.lastname: Borsani
authors.forename: N
authors.initials: N
authors.lastname: Reig
authors.forename: E
authors.initials: E
authors.lastname: Fernández
authors.forename: A
authors.initials: A
authors.lastname: Zorzano
authors.forename: J
authors.initials: J
authors.lastname: Bertran
authors.forename: M
authors.initials: M
authors.lastname: Palacín
mesh_terms: D000595:Amino Acid Sequence; D026923:Amino Acid Transport Systems, Basic; D002352:Carrier Proteins / Q000737:chemistry / Q000235:genetics* / Q000502:physiology*; D002874:Chromosome Mapping; D003404:Creatinine / Q000652:urine; D003555:Cystinuria / Q000145:classification* / Q000235:genetics* / Q000652:urine; D004252:DNA Mutational Analysis; D005091:Exons / Q000235:genetics; D006580:Genetic Carrier Screening; D005838:Genotype; D006367:HeLa Cells; D006801:Humans; D008562:Membrane Glycoproteins / Q000737:chemistry / Q000172:deficiency / Q000235:genetics* / Q000502:physiology*; D008969:Molecular Sequence Data; D020125:Mutation, Missense / Q000235:genetics*; D010641:Phenotype
chemical_list: D026923:Amino Acid Transport Systems, Basic; D002352:Carrier Proteins; D008562:Membrane Glycoproteins; C499489:SLC7A9 protein, human; D003404:Creatinine