目的探讨MAP4K1在黑色素瘤(cutaneous melanoma,CM)细胞和组织中表达及对恶性CM细胞A375迁移、侵袭的影响和机制。方法收集自2010年1月至2015年12月重庆市中医院皮肤科诊治的CM患者54例,采用GEPIA在线分析CM组织中MAP4K1的表达,实时定...目的探讨MAP4K1在黑色素瘤(cutaneous melanoma,CM)细胞和组织中表达及对恶性CM细胞A375迁移、侵袭的影响和机制。方法收集自2010年1月至2015年12月重庆市中医院皮肤科诊治的CM患者54例,采用GEPIA在线分析CM组织中MAP4K1的表达,实时定量聚合酶链法和免疫组织化学法检测CM组织中MAP4K1 m RNA和蛋白的表达水平;以MAP4K1基因表达最高的A375细胞为研究对象,采用脂质体转染法将MAP4K1抑制质粒和对照质粒转染至A375细胞;免疫蛋白印迹实验和实时定量聚合酶链法分别检测MAP4K1的基因敲除;分别通过Transwell实验检测MAP4K1敲除后对细胞侵袭的影响;荧光素酶报告实验检测MAP4K1对YAP/TAZ的调控;GEPIA验证MAP4K1与Hippo-YAP通路下游靶点的表达相关性;免疫蛋白印迹实验验证MAP4K1敲除后对Hippo-YAP信号通路的影响。结果与癌旁组织相比,MAP4K1 m RNA和蛋白在皮肤恶性CM细胞和组织中的表达上调(P<0.001);MAP4K1基因敲除A375细胞迁移和侵袭的能力明显受到抑制(P<0.001);MAP4K1敲除后YAP/TAZ的转录活性及下游的靶分子下调;MAP4K1基因敲除能抑制Hippo-YAP通路传导。结论MAP4K1在CM中表达上调,可能作为潜在的癌基因调控Hippo-YAP通路促进CM的发生发展。展开更多
In higher plants,stomatal movements represent a critical physiological process that matains cellular water homestasis while enabling photosynthetic gas exchange.Open stomata 1(OST1),a key protein kinase in the abscisi...In higher plants,stomatal movements represent a critical physiological process that matains cellular water homestasis while enabling photosynthetic gas exchange.Open stomata 1(OST1),a key protein kinase in the abscisic acid(ABA)signaling cascade,has been established as a central regulator of stomatal dynamics.This study reveals that two highly conserved mitogen-activated protein kinase 1(MAP4K1)and MAP4K2 are positive regulators in ABA promoted stomatal closure,and ABA-activated OST1 potentiates MAP4K1/2 through phosphorylation at conserved serine and threonine residues(S166,T170,and S479/S488).The activated MAP4K1,in turn,phosphorylates two critical downstream targets:plasma membrane H+-ATPase 2(AHA2)at residues T858,T881,and Y946,and slow anion channel-associated 1(SLAC1)at T114 and S116.Functional analysis demonstrates that the phosphomimetic(3D:S166D/T170D/S479D)MAP4K1,but not non-phosphorylatable(3A:S166A/T170A/S479A)MAP4K1,could fully restore drought tolerance and reduced water loss in detached leaves of map4k1map4k2 double mutant.Our findings delineate a previously unrecognized signaling module comprising OST1-MAP4K1/2-AHA2/SLAC1,which crucially modulates ABA-mediated stomatal regulation.This work advances our mechanistic understanding of phosphorylation cascades governing plant water relations and stress responses.展开更多
文摘目的探讨MAP4K1在黑色素瘤(cutaneous melanoma,CM)细胞和组织中表达及对恶性CM细胞A375迁移、侵袭的影响和机制。方法收集自2010年1月至2015年12月重庆市中医院皮肤科诊治的CM患者54例,采用GEPIA在线分析CM组织中MAP4K1的表达,实时定量聚合酶链法和免疫组织化学法检测CM组织中MAP4K1 m RNA和蛋白的表达水平;以MAP4K1基因表达最高的A375细胞为研究对象,采用脂质体转染法将MAP4K1抑制质粒和对照质粒转染至A375细胞;免疫蛋白印迹实验和实时定量聚合酶链法分别检测MAP4K1的基因敲除;分别通过Transwell实验检测MAP4K1敲除后对细胞侵袭的影响;荧光素酶报告实验检测MAP4K1对YAP/TAZ的调控;GEPIA验证MAP4K1与Hippo-YAP通路下游靶点的表达相关性;免疫蛋白印迹实验验证MAP4K1敲除后对Hippo-YAP信号通路的影响。结果与癌旁组织相比,MAP4K1 m RNA和蛋白在皮肤恶性CM细胞和组织中的表达上调(P<0.001);MAP4K1基因敲除A375细胞迁移和侵袭的能力明显受到抑制(P<0.001);MAP4K1敲除后YAP/TAZ的转录活性及下游的靶分子下调;MAP4K1基因敲除能抑制Hippo-YAP通路传导。结论MAP4K1在CM中表达上调,可能作为潜在的癌基因调控Hippo-YAP通路促进CM的发生发展。
基金supported by grants from the National Science Foundation of China(31921001)the Beijing Outstanding University Discipline。
文摘In higher plants,stomatal movements represent a critical physiological process that matains cellular water homestasis while enabling photosynthetic gas exchange.Open stomata 1(OST1),a key protein kinase in the abscisic acid(ABA)signaling cascade,has been established as a central regulator of stomatal dynamics.This study reveals that two highly conserved mitogen-activated protein kinase 1(MAP4K1)and MAP4K2 are positive regulators in ABA promoted stomatal closure,and ABA-activated OST1 potentiates MAP4K1/2 through phosphorylation at conserved serine and threonine residues(S166,T170,and S479/S488).The activated MAP4K1,in turn,phosphorylates two critical downstream targets:plasma membrane H+-ATPase 2(AHA2)at residues T858,T881,and Y946,and slow anion channel-associated 1(SLAC1)at T114 and S116.Functional analysis demonstrates that the phosphomimetic(3D:S166D/T170D/S479D)MAP4K1,but not non-phosphorylatable(3A:S166A/T170A/S479A)MAP4K1,could fully restore drought tolerance and reduced water loss in detached leaves of map4k1map4k2 double mutant.Our findings delineate a previously unrecognized signaling module comprising OST1-MAP4K1/2-AHA2/SLAC1,which crucially modulates ABA-mediated stomatal regulation.This work advances our mechanistic understanding of phosphorylation cascades governing plant water relations and stress responses.
