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Journal of Integrative Plant Biology 2023年第6期65卷 1585-1601页

作者： Yuan Li Hanwei Cao Tingting Dong Xiaoke Wang Liang Ma Kun Li Huiqiang Lou Chun-Peng Song Dongtao Ren State Key Laboratory of Plant Environmental Resilience College of Biological SciencesChina Agricultural UniversityBeijing 100193China State Key Laboratory of Agro-Biotechnology and Beijing Advanced Innovation Center for Food Nutrition and Human Health College of Biological SciencesChina Agricultural UniversityBeijing 100193China Collaborative Innovation Center of Crop Stress Biology Henan Province.Institute of Plant Stress BiologySchool of Life ScienceHenan UniversityKaifeng 475001China

Sphingolipids are the structural components of membrane lipid bilayers and act as signaling molecules in many cellular *** palmitoyltransferase(SPT)is the first committed and rate-limiting enzyme in the de novo sphingolipids biosynthetic *** core SPT enzyme is a heterodimer consisting of LONG-CHAIN BASE1(LCB1)and LCB2 *** activity is inhibited by orosomucoid proteins and stimulated by small subunits of SPT(ssSPTs).However,whether LCB1 is modified and how such modification might regulate SPT activity have to date been ***,we show that activation of MITOGEN-ACTIVATED PROTEIN KINASE 3(MPK3)and MPK6 by upstream MKK9 and treatment with Flg22(a pathogen-associated molecular pattern)increases SPT activity and induces the accumulation of sphingosine long-chain base t18:0 in Arabidopsis thaliana,with activated MPK3and MPK6 phosphorylating *** of AtLCB1 strengthened its binding with AtLCB2b,promoted its binding with ssSPTs,and stimulated the formation of higher order oligomeric and active SPT *** findings therefore suggest a novel regulatory mechanism for SPT activity.

关键词： Arabidopsis MAPK phosphorylation sphingolipids

phosphorylation of CENP-R by Aurora B regulates kinetochore-microtubule attachment for accurate chromosome segregation

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Journal of Molecular Cell Biology 2022年第7期14卷 62-75页

作者： Divine Mensah Sedzro Xiao Yuan McKay Mullen Umer Ejaz Tongtong Yang Xu Liu Xiaoyu Song Yun-Chi Tang Weijun Pan Peng Zou Xinjiao Gao Dongmei Wang Zhikai Wang Zhen Dou Xing Liu Xuebiao Yao MOE Key Laboratory for Cellular Dynamics University of Science and Technology of China School of Life SciencesHefei 230027China Anhui Key Laboratory for Cellular Dynamics and Chemical Biology National Center for Cross-Disciplinary Sciences&CAS Center for Excellence in Molecular Cell ScienceHefei 230026China Morehouse School of Medicine AtlantaGA 30310USA Shanghai Institute of Nutrition and Health Shanghai Institutes for Biological SciencesChinese Academy of Sciences(CAS)Shanghai 200031China College of Chemistry and Molecular Engineering Peking UniversityBeijing 100871China

Error-free mitosis depends on accurate chromosome attachment to spindle microtubules via a fine structure called the centromere that is epigenetically specified by the enrichment of CENP-A *** maintenance during mitosis requires CENP-A-mediated deposition of constitutive centromere-associated network that establishes the inner kinetochore and connects centromeric chromatin to spindle microtubules during *** previously proposed to be an adaptor of retinoic acid receptor,here,we show that CENP-R synergizes with CENP-OPQU to regulate kinetochore-microtubule attachment stability and ensure accurate chromosome segregation in *** found that a phospho-mimicking mutation of CENP-R weakened its localization to the kinetochore,suggesting that phosphorylation may regulate its *** of CENP-R phosphorylation is shown to prevent proper kinetochore-microtubule attachment at ***,CENP-R phosphorylation disrupts its binding with ***,we speculate that Aurora B-mediated CENP-R phosphorylation promotes the correction of improper kinetochore-microtubule attachment in *** CENP-R is absent from yeast,we reasoned that metazoan evolved an elaborate chromosome stability control machinery to ensure faithful chromosome segregation in mitosis.

关键词： constitutive centromere-associated network(CCAN) kinetochore microtubule Aurora B CENP-R phosphorylation

phosphorylation regulates cullin-based ubiquitination in tumorigenesis

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Acta Pharmaceutica Sinica B 2021年第2期11卷 309-321页

作者： Yifan Chen Xuejing Shao Ji Cao Hong Zhu Bo Yang Qiaojun He Meidan Ying Zhejiang Province Key Laboratory of Anti-Cancer Drug Research College of Pharmaceutical SciencesZhejiang UniversityHangzhou 310058China

Cullin-RING ligases(CRLs)recognize and interact with substrates for ubiquitination and degradation,and can be targeted for disease treatment when the abnormal expression of substrates involves pathologic ***,either of substrates or receptors of CRLs,can alter their ***-dependent ubiquitination and proteasome degradation influence various cellular processes and can contribute to the occurrence of various diseases,most often *** processes have the potential to be used for tumor intervention through the regulation of the activities of related kinases,along with the regulation of the stability of specific oncoproteins and tumor *** review describes the mechanisms and biological functions of crosstalk between phosphorylation and ubiquitination,and most importantly its influence on tumorigenesis,to provide new directions and strategies for tumor therapy.

关键词： phosphorylation Cullin-RING ligases Ubiquitination Tumorigenesis Targeted therapy Degradation Crosstalk Kinases

phosphorylation of a wheat aquaporin at two sites enhances both plant growth and defense

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Molecular Plant 2022年第11期15卷 1772-1789页

作者： Kai Lu Xiaochen Chen Xiaohui Yao Yuyan An Xuan Wang Lina Qin Xiaoxu Li Zuodong Wang Shuo Liu Zhimao Sun Liyuan Zhang Lei Chen Baoyan Li Baoyou Liu Weiyang Wang Xinhua Ding Yonghua Yang Meixiang Zhang Shenshen Zou Hansong Dong College of Plant Protection State Key Laboratory of Crop BiologyQilu CollegeShandong Agricultural UniversityTaian 271018China College of Life Sciences Shaanxi Normal UniversityXi’an 710019China Institute of Plant Molecular Biology State Key Laboratory of Pharmaceutical BiotechnologySchool of Life SciencesNanjing UniversityNanjing 210023China nstitute of Plant Protection&Resource and Environment Yantai Academy of Agricultural SciencesYantai 265599China

Eukaryotic aquaporins share the characteristic of functional multiplicity in transporting distinct substrates and regulating various processes,but the underlying molecular basis for this is largely ***,we report that the wheat(Triticum aestivum)aquaporin TaPIP2;10 undergoes phosphorylation to promote photosynthesis and productivity and to confer innate immunity against pathogens and a generalist aphid *** response to elevated atmospheric CO_(2)concentrations,TaPIP2;10 is phosphorylated at the serine residue S280 and thereafter transports CO_(2)into wheat cells,resulting in enhanced photosynthesis and increased grain *** response to apoplastic H_(2)O_(2) induced by pathogen or insect attacks,TaPIP2;10 is phosphorylated at S121 and this phosphorylated form transports H_(2)O_(2) into the cytoplasm,where H_(2)O_(2)intensifies host defenses,restricting further *** resistance and grain yield could be simultaneously increased by TaPIP2;10 overexpression or by expressing a TaPIP2;10 phosphomimic with aspartic acid substitutions at S121 and S280,thereby improving both crop productivity and immunity.

关键词： aquaporin phosphorylation gating photosynthesis immunity growth-defense tradeoffs

phosphorylation of human Sgo1 by NEK2A is essential for chromosome congression in mitosis

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Cell Research 2007年第7期17卷 608-618页

作者： Guosheng Fu1 Xia Ding2,3 Kai Yuan1,2 Felix Aikhionbare2 Jianhui Yao1 Xin Cai1,2 Kai Jiang1,2 Xuebiao Yao1,21 Laboratory of Cellular Dynamics, Hefei National Laboratory, University of Science & Technology of China, Hefei 230027, China2 Department of Physiology and Cancer Biology Program, Morehouse School of Medicine, Atlanta, GA 30310, USA3 Department of Medicine, Beijing University of Chinese Medicine, Beijing 100029, China Laboratory of Cellular Dynamics Hefei National Laboratory University of Science & Technology of China Hefei China Department of Physiology and Cancer Biology Program Morehouse School of Medicine Atlanta USA Department of Medicine Beijing University of Chinese Medicine Beijing China

Chromosome segregation in mitosis is orchestrated by the interaction of the kinetochore with spindle microtubules. Ourrecent study shows that NEK2A interacts with MAD1 at the kinetochore and possibly functions as a novel integrator ofspindle checkpoint signaling. However, it is unclear how NEK2 A regulates kinetochore-microtubule attachment in *** we show that NEK2A phosphorylates human Sgol and such phosphorylation is essential for faithful chromosomecongression in mitosis. NEK2A binds directly to HsSgol in vitro and co-distributes with HsSgol to the kinetochore ofmitotic cells. Our in vitro phosphorylation experiment demonstrated that HsSgol is a substrate of NEK2A and the phos-phorylation sites were mapped to Ser14and Ser507as judged by the incorporation of32P. Although such phosphorylation isnot required for assembly of HsSgol to the kinetochore, expression of non-phosphorylatable mutant HsSgol perturbedchromosome congression and resulted in a dramatic increase in microtubule attachment errors, including syntelic andmonotelic attachments. These findings reveal a key role for the NEK2A-mediated phosphorylation of HsSgol in orches-trating dynamic kinetochore-microtubule interaction. We propose that NEK2 A-mediated phosphorylation of human Sgolprovides a link between centromeric cohesion and spindle microtubule attachment at the kinetochores.

关键词： Sgo1 NEK2A phosphorylation microtubule kinetochore

phosphorylation‐dependent Traffcking of Plasma Membrane Proteins in Animal and Plant Cells

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Journal of Integrative Plant Biology 2013年第9期55卷 789-808页

作者： Remko Offringa Fang Huang Molecular and Developmental Genetics Institute Biology Leiden Sylvius Laboratory Leiden University Institute of Integrative Biology University of Liverpool

In both unicellular and multicellular organisms, transmembrane （TM） proteins are sorted to and retained at specific membrane domains by endomembrane trafficking mechanisms that recognize sorting signals in the these proteins. The trafficking and distribution of plasma membrane （PM）-localized TM proteins （PM proteins）, especially of those PM proteins that show an asymmetric distribution over the PM, has received much attention, as their proper PM localization is crucial for elementary signaling and transport processes, and defects in their localization often lead to severe disease symptoms or developmental defects. The subcellular localization of PM proteins is dynamically regulated by post-translational modifications, such as phosphorylation and ubiquitination. These modificaitons mostly occur on sorting signals that are located in the larger cytosolic domains of the cargo proteins. Here we review the effects of phosphorylation of PM proteins on their trafficking, and present the key examples from the animal field that have been subject to studies for already several decades, such as that of aquaporin 2 and the epidermal growth factor receptor. Our knowledge on cargo trafficking in plants is largely based on studies of the family of PIN FORMED （PIN） carriers that mediate the efflux of the plant hormone auxin. We will review what is known on the subcellular distribution and trafficking of PIN proteins, with a focus on how this is modulated by phosphorylation, and identify and discuss analogies and differences in trafficking with the well-studied animal examples.

关键词： Ceil polarity endosomal trafficking phosphorylation PIN auxin efflux carriers plasmamembrane-iocalized transmembrane proteins.

phosphorylation of PB2 at serine 181 restricts viral replication and virulence of the highly pathogenic H5N1 avian influenza virus in mice

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Virologica Sinica 2024年第1期39卷 97-112页

作者： Jiao Hu Zixiong Zeng Xia Chen Manyu Zhang Zenglei Hu Min Gu Xiaoquan Wang Ruyi Gao Shunlin Hu Yu Chen Xiaowen Liu Daxin Peng Xiufan Liu Animal Infectious Disease Laboratory School of Veterinary MedicineYangzhou UniversityYangzhou225009China Joint International Research Laboratory of Agriculture and Agri-Product Safety The Ministry of Education of ChinaYangzhou University225009China Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonosis Yangzhou UniversityYangzhou225009China Key Laboratory of Prevention and Control of Biological Hazard Factors(Animal Origin)for Agri-food Safety and Quality Ministry of Agriculture of China(26116120)Yangzhou UniversityYangzhou225009China

Influenza A virus(IAV)continues to pose a pandemic threat to public health,resulting a high mortality rate annually and during pandemic *** modification of viral protein plays a substantial role in regulating IAV ***,based on immunoprecipitation(IP)-based mass spectrometry(MS)and purified virus-coupled MS,a total of 89 phosphorylation sites distributed among 10 encoded viral proteins of IAV were identified,including 60 novel phosphorylation ***,for the first time,we provide evidence that PB2 can also be acetylated at site ***,the PB2 S181 phosphorylation site was consistently identified in both IP-based MS and purified virus-based *** S181 and K187 are exposed on the surface of the PB2 protein and are highly conserved in various IAV strains,suggesting their fundamental importance in the IAV life *** analysis results demonstrated that S181E/A and K187Q/R mimic mutations do not significantly alter the PB2 protein *** continuous phosphorylation mimicked by the PB2 S181E mutation substantially decreases viral fitness in mice,PB2 K187Q mimetic acetylation slightly enhances viral virulence in ***,PB2 S181E substantially impairs viral polymerase activity and viral replication,remarkably dampens protein stability and nuclear accumulation of PB2,and significantly weakens IAV-induced inflammatory ***,our study further enriches the database of phosphorylation and acetylation sites of influenza viral proteins,laying a foundation for subsequent mechanistic ***,the unraveled antiviral effect of PB2 S181E mimetic phosphorylation may provide a new target for the subsequent study of antiviral drugs.

关键词： H5N1 influenza virus PB2 phosphorylation Acetylation Viral fitness Mice

phosphorylation of Microtubule-associated Protein SB401 from Solanum berthaultii Regulates Its Effect on Microtubules

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Journal of Integrative Plant Biology 2009年第3期51卷 235-242页

作者： Bao-Quan Liu Lifeng Jin Lei Zhu Jiejie Li Shuli Huang Ming Yuan State Key Laboratory of Plant Physiology and Biochemistry Department of Plant Sciences College of Biological Sciences China Agricultural University Beijing 100094 China Department of Biological Engineering College of Life Sciences Dalian Nationalities University Dalian 116600 China

We reported previously that the protein SB401 from Solanum berthaultii binds to and bundles both microtubules and F-actin. In the current study, we investigated the regulation of SB401 activity by its phosphorylation. Our experimental results showed that the phosphorylation of SB401 by casein kinase II （CKII） downregulates the activities of SB401, namely the bundling of microtubules and enhancement of the polymerization of tubulin. However, phosphorylation of SB401 had no observable effect on its bundling of F-actin. Further investigation using extract of potato pollen indicated that a CKIl-like kinase may exist in potato pollen. Antibodies against CKII alpha recognized specifically a major band from the pollen extract and the pollen extract was able to phosphorylate the SB401 protein in vitro. The CKIl-like kinase showed a similar ability to downregulate the bundling of microtubules. Our experiments demonstrated that phosphorylation plays an important role in the regulation of SB401 activity. We propose that this phosphorylation may regulate the effects of SB401 on microtubules and the actin cytoskeleton.

关键词： F-actin microtubule-associated protein microtubules phosphorylation Solanum berthaultii.

phosphorylation of OsFD1 by OsCIPK3 promotes the formation of RFT1-containing florigen activation complex for long-day flowering in rice

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Molecular Plant 2021年第7期14卷 1135-1148页

作者： Qiang Peng Chunmei Zhu Tao Liu Shuo Zhang Shijing Feng Changyin Wu National Key Laboratory of Crop Genetic Improvement National Center of Plant Gene Research(Wuhan)Huazhong Agricultural UniversityWuhan 430070China Guizhou Rice Research Institute Guizhou Academy of Agricultural SciencesGuiyang 550006China

Heading date is a critical trait that determines the regional adaptability and grain productivity of many *** rice is a facultative short-day plant,its domestication led to the Ghd7-Ehd1-Hd3a/RFT1 pathway for adaptation to long-day conditions(LDs).The formation of the"florigen activation complex"(FAC)containing florigen Hd3a has been ***,the molecular composition of the FAC that contains RFT1 for long-day flowering is *** show here that RFT1 forms a ternary FAC with 14-3-3 proteins and OsFD1 to promote flowering under *** identified a calcineurin B-like-interacting protein kinase,OsCIPK3,which directly interacts with and phosphorylates OsFD1,thereby facilitating the localization of the FAC to the *** in OsCIPK3 results in a late heading date under LDs but a normal heading date under short-day ***,our results suggest that OsCIPK3 phosphorylates OsFD1 to promote RFT1-containing FAC formation and consequently induce flowering in rice under LDs.

关键词： rice heading date florigen activation complex OsCIPK3 phosphorylation

phosphorylation of Ago2 is required for its role in DNA double-strand break repair

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Journal of Genetics and Genomics 2021年第4期48卷 333-340页

作者： Xiaolu Hu Yan Li Tianfang Zhang Lin Li She Chen Xiaohong Wu Haijun Li Binjie Qi Zuobing Chen Department of Rehabilitation Medicine The First Affiliated HospitalCollege of MedicineZhejiang UniversityHangzhou 310000China Center for Plant Biology School of Life SciencesTsinghua UniversityBeijing 100084China Tsinghua-Peking Center for Life Sciences Beijing 100084China National Institute of Biological Sciences Zhongguancun Life Science ParkBeijing 102206China

Repair of DNA double-strand break(DSB)is critical for the maintenance of genome integrity.A class of DSB-induced small RNAs(di RNAs)has been shown to play an important role in DSB *** humans,di RNAs are associated with Ago2 and guide the recruitment of Rad51 to DSB sites to facilitate repair by homologous recombination(HR).Ago2 activity has been reported to be regulated by phosphorylation under normal and hypoxic ***,the role of Ago2 phosphorylation in DNA damage repair is ***,we show that S672,S828,T830,and S831 of human Ago2 are phosphorylated in response to ionizing radiation(IR).S672 A mutation of Ago2 leads to significant reduction in Rad51 foci formation and HR *** further show that defective association of Ago2 S672 A variant with DSB sites,instead of defects in di RNA and Rad51 binding,may account for decreased Rad51 foci formation and HR *** study reveals a novel regulatory mechanism for the function of Ago2 in DNA repair.

关键词： Ago2 diRNA DNA repair phosphorylation Small RNA