Disease-causing mutations associated with four bestrophinopathies exhibit disparate effects on the localization, but not the oligomerization, of Bestrophin-1.
Disease-causing mutations associated with four bestrophinopathies exhibit disparate effects on the localization, but not the oligomerization, of Bestrophin-1.
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DOI：
10.1016/j.exer.2014.02.006
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发表时间：
2014-04
影响因子：
3.4
通讯作者：
Marmorstein, Alan D.
中科院分区：
文献类型：
--
作者：
Johnson, Adiv A.;Lee, Yong-Suk;Chadburn, Andrew J.;Tammaro, Paolo;Manson, Forbes D.;Marmorstein, Lihua Y.;Marmorstein, Alan D.
关键词：
BEST 1编码Bestrophin-1（Best 1），这是一种定位于视网膜色素上皮基底外侧质膜的同源寡聚整合膜蛋白。BEST 1的突变导致五种不同的视网膜变性疾病，包括成人卵黄状黄斑营养不良（AVMD）、常染色体隐性雌激素样蛋白病（ARB）、常染色体显性玻璃体视网膜脉络膜病（ADVIRC）和视网膜色素变性（RP）。这些疾病的潜在机制以及为什么突变会导致一种疾病而不是另一种疾病，在大多数情况下，都是未知的。为了深入了解这四种疾病，我们表达了28个Best 1突变体融合YFP在极化MDCK单层，并通过共聚焦显微镜和免疫荧光，活细胞FRET，和互惠免疫共沉淀实验，筛选这些突变体的缺陷定位和寡聚化。所有28个突变体都表现出与WT Best 1相当的FRET效率，并与WT Best 1共免疫沉淀，表明寡聚化未受损。RP和ADVIRC相关突变体被正确定位于细胞的基底侧质膜，而两个AVMD和大多数ARB突变体被错误定位。当共表达时，所有错误定位的突变体导致WT Best 1错误定位到细胞内区室。我们目前和过去的研究结果表明，Best 1的错误定位不是任何个体bestrophinopathy的绝对特征，发生在AVMD，BVMD和ARB中。此外，一些不引起显性疾病的ARB突变体引起Best 1的错误定位，这表明错误定位不是疾病的原因，并且可以耐受质膜上没有Best 1活性。最后，我们发现ARB截短突变体L174 Qfs *57和R200 X可以与WT Best 1形成寡聚体，表明Best 1的前174个氨基酸足以发生寡聚化。
BEST1 encodes Bestrophin-1 (Best1), a homo-oligomeric, integral membrane protein localized to the basolateral plasma membrane of the retinal pigment epithelium. Mutations in BEST1 cause five distinct retinal degenerative diseases, including adult vitelliform macular dystrophy (AVMD), autosomal recessive bestrophinopathy (ARB), autosomal dominant vitreoretinochoroidopathy (ADVIRC), and retinitis pigmentosa (RP). The mechanisms underlying these diseases and why mutations cause one disease over another are, for the most part, unknown. To gain insights into these four diseases, we expressed 28 Best1 mutants fused to YFP in polarized MDCK monolayers and, via confocal microscopy and immunofluorescence, live-cell FRET, and reciprocal co-immunoprecipitation experiments, screened these mutants for defects in localization and oligomerization. All 28 mutants exhibited comparable FRET efficiencies to and co-immunoprecipitated with WT Best1, indicating unimpaired oligomerization. RP- and ADVIRC-associated mutants were properly localized to the basolateral plasma membrane of cells, while two AVMD and most ARB mutants were mislocalized. When co-expressed, all mislocalized mutants caused mislocalization of WT Best1to intracellular compartments. Our current and past results indicate that mislocalization of Best1 is not an absolute feature of any individual bestrophinopathy, occurring in AVMD, BVMD, and ARB. Furthermore, some ARB mutants that do not also cause dominant disease cause mislocalization of Best1, indicating that mislocalization is not a cause of disease, and that absence of Best1 activity from the plasma membrane is tolerated. Lastly, we find that the ARB truncation mutants L174Qfs*57 and R200X can form oligomers with WT Best1, indicating that the first ~174 amino acids of Best1 are sufficient for oligomerization to occur.
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影响因子：
3.8
作者：
Marmorstein, Lihua Y;Wu, Jiang;McLaughlin, Precious;Yocom, John;Karl, Mike O;Neussert, Rudgar;Wimmers, Soenke;Stanton, J Brett;Gregg, Ronald G;Strauss, Olaf;Peachey, Neal S;Marmorstein, Alan D
通讯作者：
Marmorstein, Alan D
DOI：
10.1007/s00417-009-1091-9
发表时间：
2009-08
期刊：
Graefe's archive for clinical and experimental ophthalmology = Albrecht von Graefes Archiv fur klinische und experimentelle Ophthalmologie
影响因子：
--
作者：
Gouras P;Braun K;Ivert L;Neuringer M;Mattison JA
通讯作者：
Mattison JA
影响因子：
5.6
作者：
Doumanov JA;Zeitz C;Dominguez Gimenez P;Audo I;Krishna A;Alfano G;Diaz ML;Moskova-Doumanova V;Lancelot ME;Sahel JA;Nandrot EF;Bhattacharya SS
通讯作者：
Bhattacharya SS
影响因子：
17.8
作者：
Marmorstein, Alan D.;Cross, Harold E.;Peachey, Neal S.
通讯作者：
Peachey, Neal S.
影响因子：
4
作者：
Burgess, R.;MacLaren, R. E.;Manson, F. D. C.
通讯作者：
Manson, F. D. C.