Jasmonates(JAs)are plant hormones with crucial roles in development and stress resilience.They activate MYC transcription factors by mediating the proteolysis of MYC inhibitors called JAZ proteins.In the absence of JA...Jasmonates(JAs)are plant hormones with crucial roles in development and stress resilience.They activate MYC transcription factors by mediating the proteolysis of MYC inhibitors called JAZ proteins.In the absence of JA,JAZ proteins bind and inhibit MYC through the assembly of MYC–JAZ–Novel Interactor of JAZ(NINJA)–TPL repressor complexes.However,JAZ and NINJA are predicted to be largely intrinsically unstructured,which has precluded their experimental structure determination.Through a combination of biochemical,mutational,and biophysical analyses and AlphaFold-derived ColabFold modeling,we characterized JAZ–JAZ and JAZ–NINJA interactions and generated models with detailed,high-confidence domain interfaces.We demonstrate that JAZ,NINJA,and MYC interface domains are dynamic in isolation and become stabilized in a stepwise order upon complex assembly.By contrast,most JAZ and NINJA regions outside of the interfaces remain highly dynamic and cannot be modeled in a single conformation.Our data indicate that the small JAZ Zinc finger expressed in Inflorescence Meristem(ZIM)motif mediates JAZ–JAZ and JAZ–NINJA interactions through separate surfaces,and our data further suggest that NINJA modulates JAZ dimerization.This study advances our understanding of JA signaling by providing insights into the dynamics,interactions,and structure of the JAZ–NINJA core of the JA repressor complex.展开更多
基金supported by the Van Andel Institute(to K.M.)the National Science Foundation(NSF+6 种基金MCB-1922846 to K.M.)the Six Talent Peaks Project in Jiangsu Province(NY-035 to F.Z.)the Fok Ying Tong Education Foundation(161022 to F.Z.)the National Institutes of Health(grant R01 GM57795 to G.A.H.)the Chemical Sciences,Geosciences,and Biosciences Division,Basic Energy Sciences,Office of Science at the U.S.Department of Energy(grant DE–FG02–91ER20021 to G.A.H.for infrastructure support)the Michigan State University Plant Resilience Institute(for support of L.V.-C.)the Michigan AgBioResearch Project(grant MICL02278 to G.A.H.).
文摘Jasmonates(JAs)are plant hormones with crucial roles in development and stress resilience.They activate MYC transcription factors by mediating the proteolysis of MYC inhibitors called JAZ proteins.In the absence of JA,JAZ proteins bind and inhibit MYC through the assembly of MYC–JAZ–Novel Interactor of JAZ(NINJA)–TPL repressor complexes.However,JAZ and NINJA are predicted to be largely intrinsically unstructured,which has precluded their experimental structure determination.Through a combination of biochemical,mutational,and biophysical analyses and AlphaFold-derived ColabFold modeling,we characterized JAZ–JAZ and JAZ–NINJA interactions and generated models with detailed,high-confidence domain interfaces.We demonstrate that JAZ,NINJA,and MYC interface domains are dynamic in isolation and become stabilized in a stepwise order upon complex assembly.By contrast,most JAZ and NINJA regions outside of the interfaces remain highly dynamic and cannot be modeled in a single conformation.Our data indicate that the small JAZ Zinc finger expressed in Inflorescence Meristem(ZIM)motif mediates JAZ–JAZ and JAZ–NINJA interactions through separate surfaces,and our data further suggest that NINJA modulates JAZ dimerization.This study advances our understanding of JA signaling by providing insights into the dynamics,interactions,and structure of the JAZ–NINJA core of the JA repressor complex.