TY - JOUR
T1 - Mutations in autosomal dominant polycystic kidney disease 2 gene
T2 - Reduced expression of PKD2 protein in lymphoblastoid cells
AU - Aguiari, G.
AU - Manzati, E.
AU - Penolazzi, L.
AU - Micheletti, F.
AU - Augello, G.
AU - Vitali, E. D.P.
AU - Cappelli, G.
AU - Cai, Y.
AU - Reynolds, D.
AU - Somlo, S.
AU - Piva, R.
AU - Del Senno, L.
N1 - Funding Information:
Supported in part by the Associazione Italiana per la Ricerea jul Cancro, Milano; Consiglo Nazionale delle Ricerche; Progetto finalizzato; Applicazioni Cliniche della Ricerca Oncologica; Ingegneria Genetica; Ministero dell' Universită e della Ricerca Scientifica e Tecnologica, Roma; and Regione Emilia Romagna, Bologna, Italy, and Regione Emilia Romagna. G. Aguiari and G. Augello were supported in part by Cassa di Risparmio and Azienda Ospedale, Ferrara, Italy.
PY - 1999
Y1 - 1999
N2 - The polycystic kidney disease 2 (PKD2) gene, encoding a 968-amino acid integral membrane protein with six predicted membrane-spanning domains and intracellular NH2 and COOH termini, is mutated in approximately 15% of the cases of autosomal dominant polycystic kidney disease (ADPKD), a common genetic disease frequently resulting in renal failure. For a better understanding of the cause of this disorder, we searched for mutations in the PKD2 gene in two PKD2-linked families characterized by different clinical phenotypes. A common polymorphism, a nonsense mutation, and a frameshift mutation were found. Both mutations are predicted to produce truncated proteins of 314 and 386 amino acids, arrested at the first extracellular loop of the protein. Restriction enzyme analysis of polymerase chain reaction (PCR) and reverse transcriptase (RT)-PCR products, respectively, showed that mutations cosegregated with the disease and mutated alleles were expressed at the messenger RNA level in lymphoblastoid cell lines. However, in these cells, Western blot analysis showed only PKD2 normal protein, and it was expressed at a lower level than that found in cells without the PKD2 mutation. These findings suggest that in lymphoblastoid cells, the truncated protein product of the mutant allele may not be stable.
AB - The polycystic kidney disease 2 (PKD2) gene, encoding a 968-amino acid integral membrane protein with six predicted membrane-spanning domains and intracellular NH2 and COOH termini, is mutated in approximately 15% of the cases of autosomal dominant polycystic kidney disease (ADPKD), a common genetic disease frequently resulting in renal failure. For a better understanding of the cause of this disorder, we searched for mutations in the PKD2 gene in two PKD2-linked families characterized by different clinical phenotypes. A common polymorphism, a nonsense mutation, and a frameshift mutation were found. Both mutations are predicted to produce truncated proteins of 314 and 386 amino acids, arrested at the first extracellular loop of the protein. Restriction enzyme analysis of polymerase chain reaction (PCR) and reverse transcriptase (RT)-PCR products, respectively, showed that mutations cosegregated with the disease and mutated alleles were expressed at the messenger RNA level in lymphoblastoid cell lines. However, in these cells, Western blot analysis showed only PKD2 normal protein, and it was expressed at a lower level than that found in cells without the PKD2 mutation. These findings suggest that in lymphoblastoid cells, the truncated protein product of the mutant allele may not be stable.
KW - ADPKD
KW - PKD2 mutations
KW - Polycystin-2 expression
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U2 - 10.1016/S0272-6386(99)70420-8
DO - 10.1016/S0272-6386(99)70420-8
M3 - Article
C2 - 10213643
AN - SCOPUS:0032905235
SN - 0272-6386
VL - 33
SP - 880
EP - 885
JO - American Journal of Kidney Diseases
JF - American Journal of Kidney Diseases
IS - 5
ER -