Sequestosome 1 Mutations in Paget's Disease of Bone in Australia: Prevalence, Genotype/Phenotype Correlation, and a Novel Non-UBA Domain Mutation (P364S) Associated With Increased NF-κB Signaling Without Loss of Ubiquitin Binding
Sequestosome 1 Mutations in Paget's Disease of Bone in Australia: Prevalence, Genotype/Phenotype Correlation, and a Novel Non-UBA Domain Mutation (P364S) Associated With Increased NF-κB Signaling Without Loss of Ubiquitin Binding
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DOI：
10.1359/jbmr.090214
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发表时间：
2009-07-01
影响因子：
6.2
通讯作者：
Ratajczak, Thomas
中科院分区：
文献类型：
--
作者：
Rea, Sarah L.;Walsh, John P.;Ratajczak, Thomas
先前报道的与佩吉特骨病（PDB）相关的Sequestosome 1（SQSTM 1）/p62基因突变聚集在泛素相关（乌巴）结构域中或导致其缺失。本研究的目的是研究SQSTM 1突变在澳大利亚患者中的患病率、基因型/表型相关性以及位于乌巴上游的新点突变（P364 S）的功能后果。用PDB法对49例心肌细胞进行SQSTM 1基因突变筛查。此外，194例明显散发性PDB受试者通过限制性内切酶消化筛选常见的P392 L突变。将稳定表达RANK的HEK 293细胞与SQSTM 1（野生型或突变体）或空载体的表达质粒和NF-κ B荧光素酶报告基因共转染。GST-SQSTM 1（野生型和突变体）蛋白用于下拉测定，以比较单乌比喹汀结合能力。我们在筛查的49个家庭中发现了12个SQSTM 1突变（24.5%），包括9个P392 L突变家庭和1个P392 L突变家庭。该家族各有以下突变：K378 X、390 X和乌巴上游第7外显子的新P364 S突变。在194例散发性疾病患者中，9例（4.6%）发现P392 L突变。SQSTM 1受试者。与无突变的受试者相比，突变的受试者疾病范围更广，但发病时间不早。在功能研究中，与野生型SQSTM 1相比，P364 S突变增加NF-κ B活化，但不减少泛素结合。这表明NF-κ B信号的增加，而不是泛素结合的损伤，可能是与SQSTM 1突变相关的PD B的发病机制中必不可少的。骨矿研究杂志2009;24：1216-1223。在线发表于2009年2月16日; doi：10.1359/JBMR.090214
Previously reported Sequestosome 1(SQSTM1)/p62 gene mutations associated with Paget's disease of bone (PDB) cluster in, or cause deletion of, the ubiquitin-associated (UBA) domain. The aims of this study were to examine the prevalence of SQSTM1 mutations in Australian patients, genotype/phenotype correlations and the functional consequences of a novel point mutation (P364S) located upstream of the UBA. Mutation screening of the SQSTM1 gene was conducted on 49 kindreds with PDB. In addition, 194 subjects with apparently sporadic PDB were screened for the common P392L mutation by restriction enzyme digestion. HEK293 cells stably expressing RANK were co-transfected with expression plasmids for SQSTM1 (wildtype or mutant) or empty vector and a NF-kappa B luciferase reporter gene. GST-SQSTM1 (wildtype and mutant) proteins were used in pull-down assays to compare monoubiquitin-binding ability. We identified SQSTM1 mutations in 12 of 49 families screened (24.5%), comprising 9 families with the P392L mutation and 1. family each with the following mutations: K378X, 390X, and a novel P364S mutation in exon 7, upstream of the UBA. The P392L mutation was found in 9 of 194 (4.6%) patients with sporadic disease. Subjects with SQSTM1. mutations had more extensive disease, but not earlier onset, compared with subjects without mutations. In functional studies, the P364S mutation increased NF-kappa B activation compared with wildtype SQSTM1 but did not reduce ubiquitin binding. This suggests that increased NF-kappa B signaling, but not the impairment of ubiquitin binding, may be essential in the pathogenesis of PDB associated with SQSTM1 mutations. J Bone Miner Res 2009;24:1216-1223. Published online on February 16, 2009; doi: 10.1359/JBMR.090214