目的探讨自噬相关基因Atg3和自噬对非小细胞肺癌A549细胞迁移的影响及相关机制。方法饥饿处理诱导非小细胞肺癌A549细胞自噬,Transwell法检测细胞迁移能力的变化,Western blot检测自噬相关基因Atg3的表达。转染Atg3 si RNA靶向沉默Atg3...目的探讨自噬相关基因Atg3和自噬对非小细胞肺癌A549细胞迁移的影响及相关机制。方法饥饿处理诱导非小细胞肺癌A549细胞自噬,Transwell法检测细胞迁移能力的变化,Western blot检测自噬相关基因Atg3的表达。转染Atg3 si RNA靶向沉默Atg3的表达,Transwell法检测细胞迁移能力的变化、流式细胞术检测细胞凋亡、Western blot检测细胞的自噬活性以及E-cadherin、β-catenin、N-cadherin、vimentin、Notch和snail的表达。结果饥饿处理可诱导非小细胞肺癌A549细胞自噬活性增强,并提高Atg3的表达,促进细胞迁移,而不影响细胞凋亡。沉默Atg3可抑制饥饿诱导的细胞自噬及迁移,而不影响细胞凋亡。此外沉默Atg3之后,E-cadherin和β-catenin的表达明显升高,而N-cadherin、vimentin、Notch和snail的表达明显降低。结论沉默Atg3可减弱自噬活性,抑制A549细胞的迁移;其机制可能与Notch/snail信号通路活性降低进而影响细胞的EMT进程有关。展开更多
Viral infection commonly induces autophagy,leading to antiviral responses or conversely,promoting viral infection or replication.In this study,using the experimental plant Nicotiana benthamiana,we demonstrated that th...Viral infection commonly induces autophagy,leading to antiviral responses or conversely,promoting viral infection or replication.In this study,using the experimental plant Nicotiana benthamiana,we demonstrated that the rice stripe virus(RSV)coat protein(CP)enhanced autophagic activity through interaction with cytosolic glyceraldehyde-3-phosphate dehydrogenase 2(GAPC2),a negative regulator of plant autophagy that binds to an autophagy key factor,autophagy-related protein 3(ATG3).Competitive pull-down and co-immunoprecipitation(Co-IP)assays showed that RSV CP activated autophagy by disrupting the interaction between GAPC2 and ATG3.An RSV CP mutant that was unable to bind GAPC2 failed to disrupt the interaction between GAPC2 and ATG3 and therefore lost its ability to induce autophagy.RSV CP enhanced the autophagic degradation of a viral movement protein(MP)encoded by a heterologous virus,citrus leaf blotch virus(CLBV).However,the autophagic degradation of RSV-encoded MP and RNA-silencing suppressor(NS3)proteins was inhibited in the presence of CP,suggesting that RSV CP can protect MP and NS3 against autophagic degradation.Moreover,in the presence of MP,RSV CP could induce the autophagic degradation of a remorin protein(NbREM1),which negatively regulates RSV infection through the inhibition of viral cell-to-cell movement.Overall,our results suggest that RSV CP induces a selective autophagy to suppress the antiviral factors while protecting RSV-encoded viral proteins against autophagic degradation through an as-yet-unknown mechanism.This study showed that RSV CP plays dual roles in the autophagy-related interaction between plants and viruses.展开更多
自噬相关基因3（autophagy related gene 3,ATG3）及其对应的蛋白是一种泛素样缀合酶。研究发现,细胞自噬体的形成,离不开2个必需系统的参与：1个是对于ATG12与ATG5两种自噬蛋白结合形成微管相关蛋白-轻链3（microtubule-associated pro...自噬相关基因3（autophagy related gene 3,ATG3）及其对应的蛋白是一种泛素样缀合酶。研究发现,细胞自噬体的形成,离不开2个必需系统的参与：1个是对于ATG12与ATG5两种自噬蛋白结合形成微管相关蛋白-轻链3（microtubule-associated protein light chain 3,LC3）前体的调节系统,此阶段需要El样酶ATG7和E2样酶ATG10共同参与活化,1个是对于自噬体LC3蛋白的脂化系统[1],它们都属于泛素样蛋白结合系统，而ATG3作为泛素样缀合酶，在LC3蛋白的脂化系统中起到重要作用。展开更多
文摘目的探讨自噬相关基因Atg3和自噬对非小细胞肺癌A549细胞迁移的影响及相关机制。方法饥饿处理诱导非小细胞肺癌A549细胞自噬,Transwell法检测细胞迁移能力的变化,Western blot检测自噬相关基因Atg3的表达。转染Atg3 si RNA靶向沉默Atg3的表达,Transwell法检测细胞迁移能力的变化、流式细胞术检测细胞凋亡、Western blot检测细胞的自噬活性以及E-cadherin、β-catenin、N-cadherin、vimentin、Notch和snail的表达。结果饥饿处理可诱导非小细胞肺癌A549细胞自噬活性增强,并提高Atg3的表达,促进细胞迁移,而不影响细胞凋亡。沉默Atg3可抑制饥饿诱导的细胞自噬及迁移,而不影响细胞凋亡。此外沉默Atg3之后,E-cadherin和β-catenin的表达明显升高,而N-cadherin、vimentin、Notch和snail的表达明显降低。结论沉默Atg3可减弱自噬活性,抑制A549细胞的迁移;其机制可能与Notch/snail信号通路活性降低进而影响细胞的EMT进程有关。
基金supported in part by the National Natural Science Foundation of China(32170163)to LSThe program of introducing Talents of Innovative discipline to universities(project 111)(B18042)to LSThe state key laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agroproducts(project 2021DG700024-KF202210)to HZ.
文摘Viral infection commonly induces autophagy,leading to antiviral responses or conversely,promoting viral infection or replication.In this study,using the experimental plant Nicotiana benthamiana,we demonstrated that the rice stripe virus(RSV)coat protein(CP)enhanced autophagic activity through interaction with cytosolic glyceraldehyde-3-phosphate dehydrogenase 2(GAPC2),a negative regulator of plant autophagy that binds to an autophagy key factor,autophagy-related protein 3(ATG3).Competitive pull-down and co-immunoprecipitation(Co-IP)assays showed that RSV CP activated autophagy by disrupting the interaction between GAPC2 and ATG3.An RSV CP mutant that was unable to bind GAPC2 failed to disrupt the interaction between GAPC2 and ATG3 and therefore lost its ability to induce autophagy.RSV CP enhanced the autophagic degradation of a viral movement protein(MP)encoded by a heterologous virus,citrus leaf blotch virus(CLBV).However,the autophagic degradation of RSV-encoded MP and RNA-silencing suppressor(NS3)proteins was inhibited in the presence of CP,suggesting that RSV CP can protect MP and NS3 against autophagic degradation.Moreover,in the presence of MP,RSV CP could induce the autophagic degradation of a remorin protein(NbREM1),which negatively regulates RSV infection through the inhibition of viral cell-to-cell movement.Overall,our results suggest that RSV CP induces a selective autophagy to suppress the antiviral factors while protecting RSV-encoded viral proteins against autophagic degradation through an as-yet-unknown mechanism.This study showed that RSV CP plays dual roles in the autophagy-related interaction between plants and viruses.
文摘自噬相关基因3（autophagy related gene 3,ATG3）及其对应的蛋白是一种泛素样缀合酶。研究发现,细胞自噬体的形成,离不开2个必需系统的参与：1个是对于ATG12与ATG5两种自噬蛋白结合形成微管相关蛋白-轻链3（microtubule-associated protein light chain 3,LC3）前体的调节系统,此阶段需要El样酶ATG7和E2样酶ATG10共同参与活化,1个是对于自噬体LC3蛋白的脂化系统[1],它们都属于泛素样蛋白结合系统，而ATG3作为泛素样缀合酶，在LC3蛋白的脂化系统中起到重要作用。
