The crosstalk between clathrin-mediated endocytosis(CME)and the autophagy pathway has been reported in mammals;however,the interconnection of CME with autophagy has not been established in plants.Here,we report that t...The crosstalk between clathrin-mediated endocytosis(CME)and the autophagy pathway has been reported in mammals;however,the interconnection of CME with autophagy has not been established in plants.Here,we report that the Arabidopsis CLATHRIN LIGHT CHAIN(CLC)subunit 2 and 3 double mutant,clc2-1 clc3-1,phenocopies Arabidopsis AUTOPHAGY-RELATED GENE(ATG)mutants in both autoimmunity and nutrient sensitivity.Accordingly,the autophagy pathway is significantly compromised in the clc2-1 clc3-1 mutant.Interestingly,multiple assays demonstrate that CLC2 directly interacts with ATG8h/ATG8i in a domain-specific manner.As expected,both GFP-ATG8h/GFP-ATG8i and CLC2-GFP are subjected to autophagic degradation,and degradation of GFP-ATG8h is significantly reduced in the clc2-1 clc3-1 mutant.Notably,simultaneous knockout of ATG8h and ATG8i by CRISPR-Cas9 results in enhanced resistance against Golovinomyces cichoracearum,supporting the functional relevance of the CLC2-ATG8h/8i interactions.In conclusion,our results reveal a link between the function of CLCs and the autophagy pathway in Arabidopsis.展开更多
基金supported by grants from the National Natural Science Foundation of China(32170761,31571423 and 31371401 to J.-Z.L.and 91754104,31820103008,and 31670283 to J.P.)the Iowa State University Plant Sciences Institute and USDA National Institute of Food and Agriculture Hatch project 4308。
文摘The crosstalk between clathrin-mediated endocytosis(CME)and the autophagy pathway has been reported in mammals;however,the interconnection of CME with autophagy has not been established in plants.Here,we report that the Arabidopsis CLATHRIN LIGHT CHAIN(CLC)subunit 2 and 3 double mutant,clc2-1 clc3-1,phenocopies Arabidopsis AUTOPHAGY-RELATED GENE(ATG)mutants in both autoimmunity and nutrient sensitivity.Accordingly,the autophagy pathway is significantly compromised in the clc2-1 clc3-1 mutant.Interestingly,multiple assays demonstrate that CLC2 directly interacts with ATG8h/ATG8i in a domain-specific manner.As expected,both GFP-ATG8h/GFP-ATG8i and CLC2-GFP are subjected to autophagic degradation,and degradation of GFP-ATG8h is significantly reduced in the clc2-1 clc3-1 mutant.Notably,simultaneous knockout of ATG8h and ATG8i by CRISPR-Cas9 results in enhanced resistance against Golovinomyces cichoracearum,supporting the functional relevance of the CLC2-ATG8h/8i interactions.In conclusion,our results reveal a link between the function of CLCs and the autophagy pathway in Arabidopsis.
