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
The nuclear pore complex(NPC),one of the largest protein complexes in eukaryotes,serves as a physical gate to regulate nucleocytoplasmic transport.Here,we determined the 8Åresolution cryo-electron microscopic(cry...The nuclear pore complex(NPC),one of the largest protein complexes in eukaryotes,serves as a physical gate to regulate nucleocytoplasmic transport.Here,we determined the 8Åresolution cryo-electron microscopic(cryo-EM)structure of the outer rings containing nuclear ring(NR)and cytoplasmic ring(CR)from the Xenopus laevis NPC,with local resolutions reaching 4.9Å.With the aid of AlphaFold2,we managed to build a pseudoatomic model of the outer rings,including the Y complexes and flanking components.In this most comprehensive and accurate model of outer rings to date,the almost complete Y complex structure exhibits much tighter interaction in the hub region.In addition to two copies of Y complexes,each asymmetric subunit in CR contains five copies of Nup358,two copies of the Nup214 complex,two copies of Nup205 and one copy of newly identified Nup93,while that in NR contains one copy of Nup205,one copy of ELYS and one copy of Nup93.These in-depth structural features represent a great advance in understanding the assembly of NPCs.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.
基金Ministry of Science and Technology of China(2017YFA0504700 to FS and 2016YFA0500201 to CMZ)the Strategic Priority Research Program of the Chinese Academy of Sciences(XDB 37040102 to FS),and National Natural Science Foundation of China(31830020 to FS,31520103906 to CMZ)+2 种基金This work was also supported by grants from the National Science Fund for Distinguished Young Scholars(31925026 to FS)National Natural Science Foundation of China(31430051 to CMZ)National Key Research and Development Program of China(2016YFA0100501 to CMZ and 2018YFA0901102 to YZ).
文摘The nuclear pore complex(NPC),one of the largest protein complexes in eukaryotes,serves as a physical gate to regulate nucleocytoplasmic transport.Here,we determined the 8Åresolution cryo-electron microscopic(cryo-EM)structure of the outer rings containing nuclear ring(NR)and cytoplasmic ring(CR)from the Xenopus laevis NPC,with local resolutions reaching 4.9Å.With the aid of AlphaFold2,we managed to build a pseudoatomic model of the outer rings,including the Y complexes and flanking components.In this most comprehensive and accurate model of outer rings to date,the almost complete Y complex structure exhibits much tighter interaction in the hub region.In addition to two copies of Y complexes,each asymmetric subunit in CR contains five copies of Nup358,two copies of the Nup214 complex,two copies of Nup205 and one copy of newly identified Nup93,while that in NR contains one copy of Nup205,one copy of ELYS and one copy of Nup93.These in-depth structural features represent a great advance in understanding the assembly of NPCs.
