Novel nephronophthisis-associated variants reveal functional importance of MAPKBP1 dimerization for centriolar recruitment.
Novel nephronophthisis-associated variants reveal functional importance of MAPKBP1 dimerization for centriolar recruitment.
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DOI：
10.1016/j.kint.2020.05.027
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发表时间：
2020-10
影响因子：
19.6
通讯作者：
Halbritter J
中科院分区：
文献类型：
--
作者：
Schönauer R;Jin W;Ertel A;Nemitz-Kliemchen M;Panitz N;Hantmann E;Seidel A;Braun DA;Shril S;Hansen M;Shahzad K;Sandford R;Saunier S;Benmerah A;Bergmann C;Hildebrandt F;Halbritter J
MAPKBP 1的双等位基因突变最近与迟发性纤毛非依赖性肾单位结核相关。MAPKBP 1被发现在有丝分裂纺锤体极，但不能检测到初级纤毛或中心体。在这里，通过鉴定和表征新的MAPKBP 1变体，我们旨在进一步研究其在健康和疾病中的作用。通过外显子组测序、纯合性作图和靶向肾脏基因面板进行遗传分析，同时使用免疫共沉淀来探索野生型和突变蛋白质-蛋白质相互作用。MAPKBP 1在非纤毛HeLa细胞和纤毛内髓集合管细胞中的表达使得能够通过荧光显微镜进行共定位研究。通过下一代测序，我们在肾单位结核相关慢性肾脏疾病患者中鉴定了两种新的纯合MAPKBP 1剪接位点变异体。剪接位点分析显示C-末端卷曲螺旋结构域的截短和患者来源的缺失构建体失去了与旁系同源WDR 62同源二聚化和异源二聚化的能力。虽然野生型MAPKBP 1表现出中心体，基体，微管协会，突变蛋白失去了后者，并显示减少招聘细胞周期依赖中心粒结构。野生型和突变体蛋白质没有相互影响后，共表达排除显性负效应。因此，MAPKBP 1似乎是一种新的微管结合蛋白与细胞周期依赖性中心粒定位。其卷曲螺旋结构域的截短足以消除其二聚化并导致严重干扰的细胞内定位。阐明受损的二聚化对细胞周期调控和细胞内肾脏信号传导的影响可能为肾脏变性的常见机制提供新的见解。因此，由于临床表现较轻，MAPKBP 1相关的肾单位结核应考虑在成人患者，否则无法解释的慢性肾脏疾病。
Biallelic mutations in MAPKBP1 were recently associated with late-onset cilia-independent nephronophthisis. MAPKBP1 was found at mitotic spindle poles but could not be detected at primary cilia or centrosomes. Here, by identification and characterization of novel MAPKBP1 variants, we aimed at further investigating its role in health and disease. Genetic analysis was done by exome sequencing, homozygosity mapping, and a targeted kidney gene panel while coimmunoprecipitation was used to explore wild-type and mutant protein-protein interactions. Expression of MAPKBP1 in non-ciliated HeLa and ciliated inner medullary collecting duct cells enabled colocalization studies by fluorescence microscopy. By next generation sequencing, we identified two novel homozygous MAPKBP1 splice-site variants in patients with nephronophthisis-related chronic kidney disease. Splice-site analyses revealed truncation of C-terminal coiled-coil domains and patient-derived deletion constructs lost their ability to homodimerize and heterodimerize with paralogous WDR62. While wild-type MAPKBP1 exhibited centrosomal, basal body, and microtubule association, mutant proteins lost the latter and showed reduced recruitment to cell cycle dependent centriolar structures. Wild-type and mutant proteins had no reciprocal influence upon co-expression excluding dominant negative effects. Thus, MAPKBP1 appears to be a novel microtubule-binding protein with cell cycle dependent centriolar localization. Truncation of its coiled-coil domain is enough to abrogate its dimerization and results in severely disturbed intracellular localizations. Delineating the impact of impaired dimerization on cell cycle regulation and intracellular kidney signaling may provide new insights into common mechanisms of kidney degeneration. Thus, due to milder clinical presentation, MAPKBP1-associated nephronophthisis should be considered in adult patients with otherwise unexplained chronic kidney disease.
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影响因子：
64.5
作者：
Chaki M;Airik R;Ghosh AK;Giles RH;Chen R;Slaats GG;Wang H;Hurd TW;Zhou W;Cluckey A;Gee HY;Ramaswami G;Hong CJ;Hamilton BA;Cervenka I;Ganji RS;Bryja V;Arts HH;van Reeuwijk J;Oud MM;Letteboer SJ;Roepman R;Husson H;Ibraghimov-Beskrovnaya O;Yasunaga T;Walz G;Eley L;Sayer JA;Schermer B;Liebau MC;Benzing T;Le Corre S;Drummond I;Janssen S;Allen SJ;Natarajan S;O'Toole JF;Attanasio M;Saunier S;Antignac C;Koenekoop RK;Ren H;Lopez I;Nayir A;Stoetzel C;Dollfus H;Massoudi R;Gleeson JG;Andreoli SP;Doherty DG;Lindstrad A;Golzio C;Katsanis N;Pape L;Abboud EB;Al-Rajhi AA;Lewis RA;Omran H;Lee EY;Wang S;Sekiguchi JM;Saunders R;Johnson CA;Garner E;Vanselow K;Andersen JS;Shlomai J;Nurnberg G;Nurnberg P;Levy S;Smogorzewska A;Otto EA;Hildebrandt F
通讯作者：
Hildebrandt F
影响因子：
4
作者：
Kaplan, Anna;Reiner, Orly
通讯作者：
Reiner, Orly
影响因子：
16.6
作者：
Veleri, Shobi;Manjunath, Souparnika H.;Fariss, Robert N.;May-Simera, Helen;Brooks, Matthew;Foskett, Trevor A.;Gao, Chun;Longo, Teresa A.;Liu, Pinghu;Nagashima, Kunio;Rachel, Rivka A.;Li, Tiansen;Dong, Lijin;Swaroop, Anand
通讯作者：
Swaroop, Anand
影响因子：
4.6
作者：
Sgourdou P;Mishra-Gorur K;Saotome I;Henagariu O;Tuysuz B;Campos C;Ishigame K;Giannikou K;Quon JL;Sestan N;Caglayan AO;Gunel M;Louvi A
通讯作者：
Louvi A
影响因子：
9.8
作者：
Macia, Maxence S.;Halbritter, Jan;Benmerah, Alexandre
通讯作者：
Benmerah, Alexandre