Certain amino acids changes in the human Na^(+)/K^(+)-ATPase pump,ATPase Na^(+)/K^(+)transporting subunit alpha 1(ATP1A1),cause Charcot-Marie-Tooth disease type 2(CMT2)disease and refractory seizures.To develop in viv...Certain amino acids changes in the human Na^(+)/K^(+)-ATPase pump,ATPase Na^(+)/K^(+)transporting subunit alpha 1(ATP1A1),cause Charcot-Marie-Tooth disease type 2(CMT2)disease and refractory seizures.To develop in vivo models to study the role of Na^(+)/K^(+)-ATPase in these diseases,we modified the Drosophila gene homolog,Atpα,to mimic the human ATP1A1 gene mutations that cause CMT2.Mutations located within the helical linker region of human ATP1A1(I592T,A597T,P600T,and D601F)were simultaneously introduced into endogenous Drosophila Atpαby CRISPR/Cas9-mediated genome editing,generating the Atpα^(TTTF)model.In addition,the same strategy was used to generate the corresponding single point mutations in flies(Atpα^(I571T),Atpα^(A576T),Atpα^(P579T),and Atpα^(D580F)).Moreover,a deletion mutation(Atpα^(mut))that causes premature termination of translation was generated as a positive control.Of these alleles,we found two that could be maintained as homozygotes(Atpα^(I571T)and Atpα^(P579T)).Three alleles(Atpα^(A576T),Atpα^(P579)and Atpα^(D580F))can form heterozygotes with the Atpαmut allele.We found that the Atpαallele carrying these CMT2-associated mutations showed differential phenotypes in Drosophila.Flies heterozygous for Atpα^(TTTF)mutations have motor performance defects,a reduced lifespan,seizures,and an abnormal neuronal morphology.These Drosophila models will provide a new platform for studying the function and regulation of the sodium-potassium pump.展开更多
植物细胞质雄性不育(cytoplasmic male sterility,CMS)与线粒体ATP合成酶亚基基因功能异常有密切关系。为了揭示烟草atp2基因在其生长发育过程中的功能,本研究利用在线软件分析了烟草ATP2蛋白的组成结构和理化性质,绘制了atp2基因和ATP...植物细胞质雄性不育(cytoplasmic male sterility,CMS)与线粒体ATP合成酶亚基基因功能异常有密切关系。为了揭示烟草atp2基因在其生长发育过程中的功能,本研究利用在线软件分析了烟草ATP2蛋白的组成结构和理化性质,绘制了atp2基因和ATP2蛋白的分子进化树,通过在烟草中瞬时表达atp2-gfp基因明确其蛋白的亚细胞定位。结果表明,ATP2蛋白全长包含560个氨基酸,等电点6.05,相对分子质量59.7 kD,氨基酸组成中甘氨酸(Gly)、丙氨酸(Ala)、亮氨酸(Leu)含量最高;其二级结构中,螺旋、折叠片、环状结构分别占比为37.95%、14.03%、48.02%;三级结构预测表明第1~51位氨基酸是线粒体信号肽序列;不同高等植物atp2基因序列一致性达到85.7%,其ATP2蛋白序列C端高度保守,但是N端信号肽序列差异显著。分子进化分析表明,atp2基因同义(dS)替换速率显著高于非同义(dN)替换速率,且不同植物中的非同义替换速率差异显著。亚细胞定位结果表明,烟草ATP2蛋白定位于线粒体内,叶绿体内未发现其定位信号。本研究结果为后续深入探讨烟草雄性生殖发育过程中atp2的基因功能提供了理论依据。展开更多
发育性和癫痫性脑病93(Developmental and epileptic encephalopathy 93,DEE93)(MIM:618012)是一种常染色体显性遗传神经系统疾病,主要特点为精神运动发育迟缓、早发难治性癫痫和智力低下等,且与ATP6V1A基因变异有关^([1])。DEE93临床...发育性和癫痫性脑病93(Developmental and epileptic encephalopathy 93,DEE93)(MIM:618012)是一种常染色体显性遗传神经系统疾病,主要特点为精神运动发育迟缓、早发难治性癫痫和智力低下等,且与ATP6V1A基因变异有关^([1])。DEE93临床表现差异大,病情轻重不一,重者可因快速进展的早期致命性脑病而死亡,轻者仅表现为轻度至中度智力障碍,伴或不伴癫痫发作^([2-3])。ATP6V1A基因(MIM#607027)染色体位置为3q13.3,包含15个外显子,编码617个氨基酸,是组成ATP酶(V-ATPase)的一个亚基,参与介导真核细胞内的质子运输过程和细胞器的酸化。本文报告了1例全外显子测序发现ATP6V1A基因杂合致病性变异c.937G>C:p.Ala313Pro的婴儿,描述了其临床表现并复习相关文献内容,扩展了ATP6V1A基因突变相关DEE93的临床特征和遗传谱,为该病的诊断、治疗和遗传咨询提供参考。展开更多
基金supported by the Natural Science Foundation of Fujian Province,No.2020J02027the National Natural Science Foundation of China,No.31970461the Foundation of NHC Key Laboratory of Technical Evaluation of Fertility Regulation for Non-human Primate,Fujian Maternity and Child Health Hospital,No.2022-NHP-05(all to WC).
文摘Certain amino acids changes in the human Na^(+)/K^(+)-ATPase pump,ATPase Na^(+)/K^(+)transporting subunit alpha 1(ATP1A1),cause Charcot-Marie-Tooth disease type 2(CMT2)disease and refractory seizures.To develop in vivo models to study the role of Na^(+)/K^(+)-ATPase in these diseases,we modified the Drosophila gene homolog,Atpα,to mimic the human ATP1A1 gene mutations that cause CMT2.Mutations located within the helical linker region of human ATP1A1(I592T,A597T,P600T,and D601F)were simultaneously introduced into endogenous Drosophila Atpαby CRISPR/Cas9-mediated genome editing,generating the Atpα^(TTTF)model.In addition,the same strategy was used to generate the corresponding single point mutations in flies(Atpα^(I571T),Atpα^(A576T),Atpα^(P579T),and Atpα^(D580F)).Moreover,a deletion mutation(Atpα^(mut))that causes premature termination of translation was generated as a positive control.Of these alleles,we found two that could be maintained as homozygotes(Atpα^(I571T)and Atpα^(P579T)).Three alleles(Atpα^(A576T),Atpα^(P579)and Atpα^(D580F))can form heterozygotes with the Atpαmut allele.We found that the Atpαallele carrying these CMT2-associated mutations showed differential phenotypes in Drosophila.Flies heterozygous for Atpα^(TTTF)mutations have motor performance defects,a reduced lifespan,seizures,and an abnormal neuronal morphology.These Drosophila models will provide a new platform for studying the function and regulation of the sodium-potassium pump.
文摘植物细胞质雄性不育(cytoplasmic male sterility,CMS)与线粒体ATP合成酶亚基基因功能异常有密切关系。为了揭示烟草atp2基因在其生长发育过程中的功能,本研究利用在线软件分析了烟草ATP2蛋白的组成结构和理化性质,绘制了atp2基因和ATP2蛋白的分子进化树,通过在烟草中瞬时表达atp2-gfp基因明确其蛋白的亚细胞定位。结果表明,ATP2蛋白全长包含560个氨基酸,等电点6.05,相对分子质量59.7 kD,氨基酸组成中甘氨酸(Gly)、丙氨酸(Ala)、亮氨酸(Leu)含量最高;其二级结构中,螺旋、折叠片、环状结构分别占比为37.95%、14.03%、48.02%;三级结构预测表明第1~51位氨基酸是线粒体信号肽序列;不同高等植物atp2基因序列一致性达到85.7%,其ATP2蛋白序列C端高度保守,但是N端信号肽序列差异显著。分子进化分析表明,atp2基因同义(dS)替换速率显著高于非同义(dN)替换速率,且不同植物中的非同义替换速率差异显著。亚细胞定位结果表明,烟草ATP2蛋白定位于线粒体内,叶绿体内未发现其定位信号。本研究结果为后续深入探讨烟草雄性生殖发育过程中atp2的基因功能提供了理论依据。
文摘发育性和癫痫性脑病93(Developmental and epileptic encephalopathy 93,DEE93)(MIM:618012)是一种常染色体显性遗传神经系统疾病,主要特点为精神运动发育迟缓、早发难治性癫痫和智力低下等,且与ATP6V1A基因变异有关^([1])。DEE93临床表现差异大,病情轻重不一,重者可因快速进展的早期致命性脑病而死亡,轻者仅表现为轻度至中度智力障碍,伴或不伴癫痫发作^([2-3])。ATP6V1A基因(MIM#607027)染色体位置为3q13.3,包含15个外显子,编码617个氨基酸,是组成ATP酶(V-ATPase)的一个亚基,参与介导真核细胞内的质子运输过程和细胞器的酸化。本文报告了1例全外显子测序发现ATP6V1A基因杂合致病性变异c.937G>C:p.Ala313Pro的婴儿,描述了其临床表现并复习相关文献内容,扩展了ATP6V1A基因突变相关DEE93的临床特征和遗传谱,为该病的诊断、治疗和遗传咨询提供参考。
