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The RING E3 ligase CLG1 targets GS3 for degradation via the endosome pathway to determine grain size in rice

Molecular Plant 2021年第10期14卷 1699-1713页

作者： Wensi Yang Kun Wu Bo Wang Huanhuan Liu Siyi Guo Xiaoyu Guo Wei Luo Shengyuan Sun Yidan Ouyang Xiangdong Fu Kang Chong Qifa Zhang Yunyuan Xu Key Laboratory of Plant Molecular Physiology Institute of BotanyChinese Academy of SciencesBeijing 100093China National Key Laboratory of Crop Genetic Improvement and National Centre of Plant Gene Research(Wuhan) Huazhong Agricultural UniversityWuhan 430070China National Key Laboratory of Crop Genetic Improvement and National Centre of Plant Gene Research(Wuhan) Hubei Hongshan LaboratoryHuazhong Agricultural UniversityWuhan 430070China University of Chinese Academy of Sciences Beijing 100049China The State Key Laboratory of Plant Cell and Chromosome Engineering Institute of Genetics and Developmental BiologyChinese Academy of SciencesBeijing 100101China Innovation Academy for Seed Design CASBeijing 100101China

G-protein signaling and ubiquitin-dependent degradation are both involved in grain development in rice,but how these pathways are coordinated in regulating this process is unknown.Here,we show that Chang Li Geng 1(CLG1),which encodes an E3 ligase,regulates grain size by targeting the Gγprotein GS3,a negative regulator of grain length,for degradation.Overexpression of CLG1 led to increased grain length,while overexpression of mutated CLG1 with changes in three conserved amino acids decreased grain length.We found that CLG1 physically interacts with and ubiquitinats GS3which is subsequently degraded through the endosome degradation pathway,leading to increased grain size.Furthermore,we identified a critical SNP in the exon 3 of CLG1 that is significantly associated with grain size variation in a core collection of cultivated rice.This SNP results in an amino acid substitution from Arg to Ser at position 163 of CLG1 that enhances the E3 ligase activity of CLG1 and thus increases rice grain size.Both the expression level of CLG1 and the SNP CLG1163S may be useful variations for manipulating grain size in rice.

关键词： grain size CLG1 GS3 E3 ligase endosome

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Adenosine Diphosphate Ribosylation Factor-GTPase-Activating Protein Stimulates the Transport of AUX1 endosome,Which Relies on Actin Cytoskeletal Organization in Rice Root Development

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Journal of Integrative Plant Biology 2011年第9期53卷 698-709页

作者： Cheng Du Yunyuan Xu Yingdian Wang Kang Chong Key Laboratory of Photosynthesis and Molecular Environmental Physiology Institute of Botanythe Chinese Academy of SciencesBeijing 100093China College of Life Sciences Beijing Normal UniversityBeijing 100875China

Polar auxin transport,which depends on polarized subcellular distribution of AUXIN RESISTANT 1/LIKE AUX1（AUX1/LAX） influx carriers and PIN-FORMED（PIN） efflux carriers,mediates various processes of plant growth and development.Endosomal recycling of PIN1 is mediated by an adenosine diphosphate（ADP）ribosylation factor（ARF）-GTPase exchange factor protein,GNOM.However,the mediation of auxin influx carrier recycling is poorly understood.Here,we report that overexpression of OsAGAP,an ARF-GTPase-activating protein in rice,stimulates vesicle transport from the plasma membrane to the Golgi apparatus in protoplasts and transgenic plants and induces the accumulation of early endosomes and AUX1.AUX1 endosomes could partially colocalize with FM4-64 labeled early endosome after actin disruption.Furthermore,OsAGAP is involved in actin cytoskeletal organization,and its overexpression tends to reduce the thickness and bundling of actin filaments.Fluorescence recovery after photobleaching analysis revealed exocytosis of the AUX1 recycling endosome was not affected in the OsAGAP overexpression cells,and was only slightly promoted when the actin filaments were completely disrupted by Lat B.Thus,we propose that AUX1 accumulation in the OsAGAP overexpression and actin disrupted cells may be due to the fact that endocytosis of the auxin influx carrier AUX1 early endosome was greatly promoted by actin cytoskeleton disruption.

关键词： actin adenosine diphosphate ribosylation factor-GTPase-activating protein AUX1 endosome polar auxin transport.

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Stay in touch with the endoplasmic reticulum

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Science China(Life Sciences) 2024年第2期67卷 230-257页

作者： Sha Sun Gan Zhao Mingkang Jia Qing Jiang Shulin Li Haibin Wang Wenjing Li Yunyun Wang Xin Bian Yan G.Zhao Xun Huang Ge Yang Huaqing Cai Jose C.Pastor-Pareja Liang Ge Chuanmao Zhang Junjie Hu National Laboratory of Biomacromolecules Institute of BiophysicsUniversity of Chinese Academy of SciencesChinese Academy of SciencesBeijing 100101China The Ministry of Education Key Laboratory of Cell Proliferation and Differentiation College of Life SciencesPeking UniversityBeijing 100871China State Key Laboratory of Membrane Biology Tsinghua-Peking Center for Life SciencesSchool of Life SciencesTsinghua UniversityBeijing 100084China Laboratory of Computational Biology&Machine Intelligence School of Artificial IntelligenceUniversity of Chinese Academy of SciencesBeijing 100049China State Key Laboratory of Medicinal Chemical Biology College of Life SciencesNankai UniversityTianjin 300071China Brain Research Center Department of BiologySchool of Life SciencesSouthern University of Science and TechnologyShenzhen 518055China State Key Laboratory of Molecular Developmental Biology Institute of Genetics and Developmental BiologyInnovation Academy for Seed DesignChinese Academy of SciencesBeijing 100101China Institute of Neurosciences Consejo Superior de Investigaciones Científicas-Universidad Miguel HernándezSan Juan de Alicante 03550Spain

The endoplasmic reticulum(ER),which is composed of a continuous network of tubules and sheets,forms the most widely distributed membrane system in eukaryotic cells.As a result,it engages a variety of organelles by establishing membrane contact sites(MCSs).These contacts regulate organelle positioning and remodeling,including fusion and fission,facilitate precise lipid exchange,and couple vital signaling events.Here,we systematically review recent advances and converging themes on ER-involved organellar contact.The molecular basis,cellular influence,and potential physiological functions for ER/nuclear envelope contacts with mitochondria,Golgi,endosomes,lysosomes,lipid droplets,autophagosomes,and plasma membrane are summarized.

关键词： endoplasmic reticulum nuclear envelope mitochondria Golgi apparatus endosome lysosome lipid droplets autophagosome plasma membrane membrane contact site

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endosomes and Microtubles are Required for Productive Infection in Aquareovirus

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Virologica Sinica 2020年第2期35卷 200-211页

作者： Fuxian Zhang Hong Guo Qingxiu Chen Zheng Ruan Qin Fang State Key Laboratory of Virology Wuhan Institute of VirologyChinese Academy of Sciences.Wuhan 430071China Wuhan Center for Animal Diseases Prevention and Control Wuhan 430071China

Grass carp reovirus(GCRV),the genus Aquareovirus in family Reoviridae,is viewed as the most pathogenic aquareovirus.To understand the molecular mechanism of how aquareovirus initiates productive infection,the roles of endosome and microtubule in cell entry of GCRV are investigated by using quantum dots(QDs)-tracking in combination with biochemical approaches.We found that GCRV infection and viral protein synthesis were significantly inhibited by pretreating host cells with endosome acidification inhibitors NH4Cl,chloroquine and bafilomycin A1(Bafi).Confocal images indicated that GCRV particles could colocalize with Rab5,Rab7 and lysosomes in host cells.Further ultrastructural examination validated that viral particle was found in late endosomes.Moreover,disruption of microtubules with nocodazole clearly blocked GCRV entry,while no inhibitory effects were observed with cytochalasin D treated cells in viral infection,hinting that intracellular transportation of endocytic uptake in GCRV infected cells is via microtubules but not actin filament.Notably,viral particles were observed to transport along microtubules by using QD-labeled GCRV.Altogether,our results suggest that GCRV can use endosomes and microtubules to initiate productive infection.

关键词： Aquareovirus Cell entry Quantum dot endosome Microtubule

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Converging links between adult-onset neurodegenerative Alzheimer’s disease and early life neurodegenerative neuronal ceroid lipofuscinosis?

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Neural Regeneration Research 2023年第7期18卷 1463-1471页

作者： Marcel Klein Guido Hermey Institute for Molecular and Cellular Cognition Center for Molecular Neurobiology HamburgUniversity Medical Center Hamburg-EppendorfHamburgGermany

Evidence from genetics and from analyzing cellular and animal models have converged to suggest links between neurodegenerative disorders of early and late life.Here,we summarize emerging links between the most common late life neurodegenerative disease,Alzheimer’s disease,and the most common early life neurodegenerative diseases,neuronal ceroid lipofuscinoses.Genetic studies reported an overlap of clinically diagnosed Alzheimer’s disease and mutations in genes known to cause neuronal ceroid lipofuscinoses.Accumulating data strongly suggest dysfunction of intracellular trafficking mechanisms and the autophagy-endolysosome system in both types of neurodegenerative disorders.This suggests shared cytopathological processes underlying these different types of neurodegenerative diseases.A better understanding of the common mechanisms underlying the different diseases is important as this might lead to the identification of novel targets for therapeutic concepts,the transfer of therapeutic strategies from one disease to the other and therapeutic approaches tailored to patients with specific mutations.Here,we review dysfunctions of the endolysosomal autophagy pathway in Alzheimer’s disease and neuronal ceroid lipofuscinoses and summarize emerging etiologic and genetic overlaps.

关键词： Alzheimer’s disease autophagy Batten disease CLN3 disease dementia endosome lysosome neurodegeneration neuronal ceroid lipofuscinosis presenilin

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Varp is a Rab21 Guanine Nucleotide Exchange Factor and Regulates endosome Dynamics

Varp is a Rab21 Guanine Nucleotide Exchange Factor and Regul...

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中国细胞生物学学会第九次会员代表大会暨青年学术大会

作者： Xin-Jun Zhang Xi He Xin-Yuan Fu Zhi-Jie Chang (School of Medicine,Tsinghua University,Beijing 100084,China)

The small GTPases Rab5 and Rab21 are closely related,and play essential roles in endocytic trafficking.Rab5 is regulated by VPS9-domain-containing guanine nucleotide exchange factors.Here,we describe a new VPS9- domai... 详细信息

关键词： ankyrin repeat endosome guanine nucleotide

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CDK11 negatively regulates Wnt/β-catenin signaling in the endosomal compartment by affecting microtubule stability

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Cancer Biology & Medicine 2020年第2期17卷 328-342页

作者： Danmin Ou Lin Chen Jiang He Zhuoxian Rong Jie Gao Liyu Liu Feiyu Tang Jiang Li Yuezhen Deng Lunquan Sun Department of Oncology Center for Molecular MedicineXiangya HospitalCentral South UniversityChangsha 410008China international Cooperation Base of Cancer Precision Therapy Department of Science and Technology of Hunan ProvinceChangsha 410008China Key Laboratory of Molecular Radiation Oncology of Hunan Province Changsha 410008China National Clinical Research Center for Geriatric Disorders Changsha 410008China

Objectives:Improper activation of Wnt/β-catenin signaling has been implicated in human diseases.Beyond the well-studied glycogen synthase kinase 3p(GSK3p)and casein kinase 1(CK1),other kinases affecting Wnt/β-catenin signaling remain to be defined.Methods:To identify the kinases that modulate Wnt/β-catenin signaling,we applied a kinase small interfering RNA(siRNA)library screen approach.Luciferase assays,immunoblotting,and real-time polymerase chain reaction(PCR)were performed to confirm the regulation o f the Wnt/β-catenin signaling pathway by cyclin-dependent kinase 11(CDK11)and to investigate the underlying mechanism.Confocal immunofluorescence,coimmunoprecipitation(co-IP),and scratch wound assays were used to demonstrate colocalization,detect protein interactions,and explore the function of CDK11.Results:CDK11 was found to be a significant candidate kinase participating in the negative control of Wnt/P-catenin signaling.Down-regulation of CDK11 led to the accumulation of Wnt/β-catenin signaling receptor complexes,in a manner dependent on intact adenomatosis polyposis coli(APC)protein.Further analysis showed that CDK11 modulation of Wnt/P-catenin signaling engaged the endolysosomal machinery,and CDK11 knockdown enhanced the colocalization of Wnt/β-catenin signaling receptor complexes with early endosomes and decreased colocalization with lysosomes.Mechanistically,CDK11 was found to function in Wnt/β-catenin signaling by regulating microtubule stability.Depletion of CDK11 down-regulated acetyl-a-tubulin.Moreover,co-IP assays demonstrated that CDK11 interacts with the a-tubulin deacetylase SIRT2,whereas SIRT2 down-regulation in CDK11-depleted cells reversed the accumulation of Wnt/(3-catenin signaling receptor complexes.CDK11 was found to suppress cell migration through altered W nt/β-catenin signaling.Conclusions:CDK11 is a negative modulator of Wnt/β-catenin signaling that stabilizes microtubules,thus resulting in the dysregulation of receptor complex trafficking from early endosomes to lysosomes.

关键词： Wnt/β-catenin signaling CDK11 endosome microtubule SIRT2

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A multi-functional nanoplatform for efficacy tumor theranostic applications

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Asian Journal of Pharmaceutical Sciences 2017年第3期12卷 235-249页

作者： Jinjin Shi Hongling Zhang Zhaoyang Chen Lihua Xu Zhenzhong Zhang School of Pharmaceutical Sciences Zhengzhou University

Nanomaterials with multiple functions have become more and more popular in the domain of cancer research. MoS2 has a great potential in photothermal therapy, X-ray/CT imaging and drug delivery. In this study, a water soluble MoS2 nanosystem(MoS2-PEG) was synthesized and explored in drug delivery, photothermal therapy(PTT) and X-ray imaging.Doxorubicin(DOX) was loaded onto MoS2-PEG with a high drug loading efficiency(~69%)and obtained a multifunctional drug delivery system(MoS2-PEG/DOX). As the drug delivery, MoS2-PEG/DOX could efficiently cross the cell membranes, and escape from the endosome via NIR light irradiation, lead to more apoptosis in MCF-7 cells, and afford higher antitumor efficacy without obvious toxic effects to normal organs owing to its prolonged blood circulation and 11.6-fold higher DTX uptake of tumor than DOX. Besides, MoS2-PEG/DOX not only served as a drug delivery system, but also as a powerful PTT agent for thermal ablation of tumor and a strong X-ray contrast agent for tumor diagnosis. In the in vitro and in vivo studies, MoS2-PEG/DOX exhibited excellent tumor-targeting efficacy, outstanding synergistic anti-cancer effect of photothermal and chemotherapy and X-ray imaging property,demonstrating that MoS2-PEG/DOX had a great potential for simultaneous diagnosis and photothermal-chemotherapy in cancer treatment.

关键词： Multifunctional drug delivery endosome escape Tumor-targeting Bio-imaging Theranostic

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Retromer-Mediated Protein Sorting and Vesicular Trafficking

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Journal of Genetics and Genomics 2016年第4期43卷 165-177页

作者： Jia-Jia Liu State Key Laboratory of Molecular Developmental Biology Institute of Genetics and Developmental BiologyChinese Academy of Sciences CAS Center for Excellence in Brain Science and Intelligence Technology Chinese Academy of Sciences

Retromer is an evolutionarily conserved multimeric protein complex that mediates intracellular transport of various vesicular cargoes and functions in a wide variety of cellular processes including polarized trafficking,developmental signaling and lysosome biogenesis.Through its interaction with the Rab GTPases and their effectors,membrane lipids,molecular motors,the endocytic machinery and actin nucleation promoting factors,retromer regulates sorting and trafficking of transmembrane proteins from endosomes to the trans-Golgi network（TGN） and the plasma membrane.In this review.I highlight recent progress in the understanding of relromer-medialed protein sorting and vesicle trafficking and discuss how retromer contributes to a diverse set of developmental,physiological and pathological processes.

关键词： Retromer Sorting nexin endosome Protein sorting and trafficking Neurodegeneration

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AtSYP51/52 Functions Diverge in the Post-Golgi Traffic and Differently Affect Vacuolar Sorting

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Molecular Plant 2013年第3期6卷 916-930页

作者： Maria De Benedictis Gianluca Bleve Marianna Faraco Egidio Stigliano Francesco Grieco Gabriella Piro Giuseppe Dalessandro Gian Pietro Di Sansebastiano Dipartimento di Scienze e Tecnologie Biologiche ed Ambientali University of Salento Campus Ecotekne 73100 Lecce Italy Present address: Dipartimento di Bioscienze Universita degli Studi di Parma Parco Area delle Scienze 11/A 43124 Parma Italy Institute of Science of Food Production National Research Council via Provinciale Lecce-Monteroni 73100 Lecce Italy Laboratory of Cell and Molecular Biology University of Neuchatel Rue Emile-Argand 11 CP 158 2009 Neuchatel Switzerland

Plant sensitive factor attachment protein receptors （SNAREs） encoded by genes of the same sub-family are generally considered as redundant in promoting vesicle-associated membrane fusion events. Nonetheless, the application of innovative experimental approaches highlighted that members of the same gene sub-family often have different functional specificities. In this work, two closely related Qc-SNAREs--the AtSYP51 and the AtSYP52--are compared in their ability to influence different secretory pathways. Their role in the vesicle sorting to the central vacuole has been revised and they were found to have a novel inhibitory function. When transiently overexpressed, the SYP51 and the SYP52 distributed between the TGN and the tonoplast. Our data demonstrate that these SYPs （syntaxin of plants） act as t-SNARE when present on the membrane of TGN/PVC, whereas they behave as inhibitory or interfering SNAREs （i-SNAREs） when they accumulate on the tonoplast. Moreover, the performed functional analysis indicated that the AtSYP51 and the AtSYP52 roles differ in the traffic to the vacuole. The findings are a novel contribution to the functional characterization of plant SNAREs that reveals additional non-fusogenic roles.

关键词： endocytosis endosome Golgi i-SNARE plant endomembranes protoplast SNARE TGN tonoplast vacuole.

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