Functional characterization of a novel GUCA1A missense mutation (D144G) in autosomal dominant cone dystrophy: A novel pathogenic GUCA1A variant in COD
Functional characterization of a novel GUCA1A missense mutation (D144G) in autosomal dominant cone dystrophy: A novel pathogenic GUCA1A variant in COD
复制标题
常染色体显性视锥细胞营养不良中新型 GUCA1A 错义突变 (D144G) 的功能特征：COD 中新型致病性 GUCA1A 变异
作者：
Tang Suzhen;Xia Yujun;Dai Yunhai;Liu Yaning;Li Jingshuo;Pan Xiaojing;Chen Peng
目的探讨鸟苷酸环化酶激活剂A1 A（GUCA 1A）基因一个新的错义突变与常染色体显性视锥细胞营养不良（adCOD）的关系。方法对一个adCOD家系成员进行临床分析。捕获相关基因，然后用靶向下一代测序进行测序，并用桑格测序进行确认。对突变残基的保守性进行了序列分析。酶联免疫吸附试验（ELISA）检测环磷酸鸟苷（cGMP）的浓度。然后使用有限的蛋白质水解和电泳位移来评估结构可能的变化。采用免疫共沉淀法分析GCAP 1和retGC 1之间的相互作用。免疫荧光染色观察GCAP 1和retGC 1在人胚肾（HEK）-293细胞中的共定位。结果在一个adCOD家系中发现了一个致病性突变位点GUCA 1A（c.431A>G，p.D144G，exon 5）。GUCA 1A编码鸟苷酸环化酶激活蛋白1（GCAP 1）。D144，位于EF 4环涉及钙结合，是高度保守的物种。GCAP 1-D144 G更容易水解，并且D144 G带的迁移率在Ca 2+存在下变得更慢。在高Ca 2+浓度下，GCAP 1-D144 G刺激HEK-293膜中的retGC 1，以显著增加细胞内cGMP蛋白浓度。与野生型（WT）GCAP 1相比，GCAP 1-D144 G与retGC 1的相互作用增加，如在免疫共沉淀试验中检测到的。结论GUCA 1A基因p.D144G突变可能导致Ca 2+和cGMP稳态失衡，导致adCOD发生显著变化。
Purpose To elucidate the clinical phenotypes and pathogenesis of a novel missense mutation in guanylate cyclase activator A1A (GUCA1A) associated with autosomal dominant cone dystrophy (adCOD). Methods The members of a family with adCOD were clinically evaluated. Relevant genes were captured before being sequenced with targeted next-generation sequencing and confirmed with Sanger sequencing. Sequence analysis was made of the conservativeness of mutant residues. An enzyme-linked immunosorbent assay (ELISA) was implemented to detect the cyclic guanosine monophosphate (cGMP) concentration. Then limited protein hydrolysis and an electrophoresis shift were used to assess possible changes in the structure. Coimmunoprecipitation was employed to analyze the interaction between GCAP1 and retGC1. Immunofluorescence staining was performed to observe the colocalization of GCAP1 and retGC1 in human embryonic kidney (HEK)-293 cells. Results A pathogenic mutation in GUCA1A (c.431A>G, p.D144G, exon 5) was revealed in four generations of a family with adCOD. GUCA1A encodes guanylate cyclase activating protein 1 (GCAP1). D144, located in the EF4 loop involving calcium binding, was highly conserved in the species. GCAP1-D144G was more susceptible to hydrolysis, and the mobility of the D144G band became slower in the presence of Ca2+. At high Ca2+ concentrations, GCAP1-D144G stimulated retGC1 in the HEK-293 membrane to significantly increase intracellular cGMP protein concentrations. Compared with wild-type (WT) GCAP1, GCAP1-D144G had an increased interaction with retGC1, as detected in the coimmunoprecipitation assay. Conclusions The newly discovered missense mutation in GUCA1A (p.D144G) might lead to an imbalance of Ca2+ and cGMP homeostasis and eventually, cause a significant variation in adCOD.
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影响因子：
2.2
作者：
G. Manes;Sonia Mamouni;E. Hérald;A. Richard;A. Sénéćhal;Karim Aouad;B. Bocquet;I. Meunier;C. Hamel
通讯作者：
G. Manes;Sonia Mamouni;E. Hérald;A. Richard;A. Sénéćhal;Karim Aouad;B. Bocquet;I. Meunier;C. Hamel
影响因子：
3.7
作者：
Lim S;Peshenko IV;Dizhoor AM;Ames JB
通讯作者：
Ames JB
影响因子：
34.7
作者：
Burgoyne, Robert D.
通讯作者：
Burgoyne, Robert D.
影响因子：
5.7
作者：
Stephen, Ricardo;Bereta, Grzegorz;Sousa, Marcelo Carlos
通讯作者：
Sousa, Marcelo Carlos
DOI：
10.1523/jneurosci.2985-17.2018
发表时间：
2018-02
期刊：
The Journal of Neuroscience
影响因子：
--
作者：
Shinya Sato;I. Peshenko;E. Olshevskaya;V. Kefalov;A. Dizhoor
通讯作者：
Shinya Sato;I. Peshenko;E. Olshevskaya;V. Kefalov;A. Dizhoor