Papillorenal syndrome-causing missense mutations in PAX2/Pax2 result in hypomorphic alleles in mouse and human.
Papillorenal syndrome-causing missense mutations in PAX2/Pax2 result in hypomorphic alleles in mouse and human.
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DOI：
10.1371/journal.pgen.1000870
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发表时间：
2010-03-05
期刊：
影响因子：
4.5
通讯作者：
Brooks BP
中科院分区：
文献类型：
--
作者：
Alur RP;Vijayasarathy C;Brown JD;Mehtani M;Onojafe IF;Sergeev YV;Boobalan E;Jones M;Tang K;Liu H;Xia CH;Gong X;Brooks BP
视神经乳头肾综合征（PRS，也称为肾缺损综合征）是一种常染色体显性遗传疾病，其特征是潜在致盲的先天性视神经凹陷和先天性肾脏异常。许多PRS患者在配对盒转录因子基因PAX 2中存在突变。虽然PAX 2中的大多数突变被预测会导致一个等位基因功能的完全丧失，但已经报道了三个错义突变，这提高了PAX 2功能中更微妙的改变可能是致病的可能性。到目前为止，这些突变的分子行为还没有被探索。我们描述了一种新的小鼠模型的PRS由于错义突变的高度保守的苏氨酸残基的Pax 2（p.T74A）的配对结构域，概括了患者的眼睛和肾脏的结果。该突变位于Pax 2配对结构域中与两个人类错义突变相同的位置。我们发现，所有三个错义突变破坏潜在的关键氢键原子模型，并导致Pax 2反式激活减少，但不影响核定位，稳态mRNA水平，或Pax 2的能力，结合其DNA的共识序列。此外，这些突变显示Pax 2蛋白在体外和（对于p.T74A）体内的稳态水平降低，可能是通过降低蛋白质稳定性。这些结果表明PAX 2/Pax 2的亚纯型等位基因可导致人类和小鼠的显著疾病。影响视神经的先天性眼部畸形是儿童失明的重要原因。视神经乳头肾综合征（PRS）是一种常染色体显性遗传疾病，可引起先天性视神经和肾脏异常，分别导致法律的失明和肾功能衰竭。许多PRS病例是由配对盒转录因子PAX 2的突变引起的。在本文中，我们描述了一种新的小鼠模型，这种人类疾病所造成的错义突变的Pax 2基因在同一位置的少数致病错义突变在人类之一。我们描述了这种小鼠的眼部和非眼部表型，并模拟了小鼠和人Pax 2/PAX 2突变对蛋白质结构的影响。我们还通过实验测试了这些错义突变对蛋白质定位、反式激活和DNA结合的影响，得出的结论是，所有这三种突变都通过降低蛋白质稳定性来降低体外和体内（在p.T74A中）蛋白质的稳态水平。这项工作将帮助我们更好地了解PRS的病理生理学，并剖析正常PAX 2功能中重要的分子相互作用。
Papillorenal syndrome (PRS, also known as renal-coloboma syndrome) is an autosomal dominant disease characterized by potentially-blinding congenital optic nerve excavation and congenital kidney abnormalities. Many patients with PRS have mutations in the paired box transcription factor gene, PAX2. Although most mutations in PAX2 are predicted to result in complete loss of one allele's function, three missense mutations have been reported, raising the possibility that more subtle alterations in PAX2 function may be disease-causing. To date, the molecular behaviors of these mutations have not been explored. We describe a novel mouse model of PRS due to a missense mutation in a highly-conserved threonine residue in the paired domain of Pax2 (p.T74A) that recapitulates the ocular and kidney findings of patients. This mutation is in the Pax2 paired domain at the same location as two human missense mutations. We show that all three missense mutations disrupt potentially critical hydrogen bonds in atomic models and result in reduced Pax2 transactivation, but do not affect nuclear localization, steady state mRNA levels, or the ability of Pax2 to bind its DNA consensus sequence. Moreover, these mutations show reduced steady-state levels of Pax2 protein in vitro and (for p.T74A) in vivo, likely by reducing protein stability. These results suggest that hypomorphic alleles of PAX2/Pax2 can lead to significant disease in humans and mice. Congenital ocular malformations affecting the optic nerve are an important cause of childhood blindness. The papillorenal syndrome (PRS) is an autosomal dominant disorder that causes congenital optic nerve and kidney abnormalities, which may result in legal blindness and renal failure, respectively. Many cases of PRS are caused by mutations in the paired-box transcription factor PAX2. In this paper, we describe a novel mouse model of this human disease caused by a missense mutation in the Pax2 gene at the same position of one of the few disease-causing missense mutations in humans. We characterize the ocular and non-ocular phenotypes of this mouse and model the effect that murine and human Pax2/PAX2 mutations have on protein structure. We also experimentally test the effect these missense mutations have on protein localization, transactivation, and DNA binding, concluding that all three reduce steady-state levels of protein in vitro and (in p.T74A) in vivo by reducing protein stability. This work will help us better understand the pathophysiology of PRS and to dissect the molecular interactions important in normal PAX2 function.
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影响因子：
5.3
作者：
Devriendt, K;Matthijs, G;Leys, A
通讯作者：
Leys, A
影响因子：
5.3
作者：
Gong, Ke-Qin;Yallowitz, Alisha R.;Wellik, Deneen M.
通讯作者：
Wellik, Deneen M.
影响因子：
5.2
作者：
D'Elia, Angela Valentina;Puppin, Cinzia;Damante, Giuseppe
通讯作者：
Damante, Giuseppe
DOI：
10.1073/pnas.93.24.13870
发表时间：
1996-11-26
影响因子：
11.1
作者：
Favor, J;Sandulache, R;Schughart, K
通讯作者：
Schughart, K
影响因子：
4.2
作者：
Higashide, T;Wada, T;Sugiyama, K
通讯作者：
Sugiyama, K