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A novel bat coronavirus with a polybasic furin-like cleavage site

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Virologica Sinica 2023年第3期38卷 344-350页

作者： Wentao Zhu Yuyuan Huang Jian Gong Lingzhi Dong Xiaojie Yu Haiyun Chen Dandan Li Libo Zhou Jing Yang Shan Lu Department of Infectious Diseases and Clinical Microbiology Beijing Institute of Respiratory Medicine and Beijing Chao-Yang HospitalCapital Medical UniversityBeijing100020China State Key Laboratory of Infectious Disease Prevention and Control National Institute for Communicable Disease Control and PreventionChinese Center for Disease Control and PreventionBeijing102206China Hainan Provincial Center for Disease Control and Prevention Haikou570203China Research Units of Discovery of Unknown Bacteria and Function Chinese Academy of Medical SciencesBeijing100730China

The current pandemic of COVID-19 caused by a novel coronavirus,severe acute respiratory syndrome coronavirus-2(SARS-CoV-2),threatens human health around the world.Of particular concern is that bats are recognized as one of the most potential natural hosts of SARS-CoV-2;however,coronavirus ecology in bats is still nascent.Here,we performed a degenerate primer screening and next-generation sequencing analysis of 112 bats,collected from Hainan Province,China.Three coronaviruses,namely bat betacoronavirus(bat CoV)CD35,bat CoV CD36 and bat alpha-coronavirus CD30 were identified.bat CoV CD35 genome had 99.5%identity with bat CoV CD36,both sharing the highest nucleotide identity with bat Hp-betacoronavirus Zhejiang2013(71.4%),followed by SARS-CoV-2(54.0%).Phylogenetic analysis indicated that bat CoV CD35 formed a distinct clade,and together with bat Hp-betacoronavirus Zhejiang2013,was basal to the lineage of SARS-CoV-1 and SARS-CoV-2.Notably,bat CoV CD35 harbored a canonical furin-like S1/S2 cleavage site that resembles the corresponding sites of SARS-CoV-2.The furin cleavage sites between CD35 and CD36 are identical.In addition,the receptor-binding domain of bat CoV CD35 showed a highly similar structure to that of SARS-CoV-1 and SARS-CoV-2,especially in one binding loop.In conclusion,this study deepens our understanding of the diversity of coronaviruses and provides clues about the natural origin of the furin cleavage site of SARS-CoV-2.

关键词： bat Coronavirus SARS-CoV-2 Betacoronavirus Furin Cleavage site

Discovery and characterization of novel paramyxoviruses from bat samples in China

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Virologica Sinica 2023年第2期38卷 198-207页

作者： Haoxiang Su Yuyang Wang Yelin Han Qi Jin Fan Yang Zhiqiang Wu NHC Key Laboratory of Systems Biology of Pathogens Institute of Pathogen BiologyChinese Academy of Medical Sciences&Peking Union Medical CollegeBeijing100730China Key Laboratory of Respiratory Disease Pathogenomics Chinese Academy of Medical Sciences&Peking Union Medical CollegeBeijing100730China

Many paramyxoviruses are responsible for a variety of mild to severe human and animal diseases.Based on the novel discoveries over the past several decades,the family Paramyxoviridae infecting various hosts across the world includes 4 subfamilies,17 classified genera and 78 species now.However,no systematic surveys of bat paramyxoviruses are available from the Chinese mainland.In this study,13,064 samples from 54 bat species were collected and a comprehensive paramyxovirus survey was conducted.We obtained 94 new genome sequences distributed across paramyxoviruses from 22 bat species in seven provinces.Bayesian phylodynamic and phylogenetic analyses showed that there were four different lineages in the Jeilongvirus genus.Based on available data,results of host and region switches showed that the bat colony was partial to interior,whereas the rodent colony was exported,and the felines and hedgehogs were most likely the intermediate hosts from Scotophilus spp.rather than rodents.Based on the evolutionary trend,genus Jeilongvirus may have originated from Mus spp.in Australia,then transmitted to bats and rodents in Africa,Asia and Europe,and finally to bats and rodents in America.

关键词： Paramyxovirus Jeilongvirus bat Host and region switches China

Genomic landscape of mitochondrial DNA insertions in 23 bat genomes:characteristics,loci,phylogeny,and polymorphism

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Integrative Zoology 2022年第5期17卷 890-903页

作者： Guojun ZHANG Deqi GENG Qiulin GUO Wei LIU Shufen LI Wujun GAO Yongfei WANG Min ZHANG Yilin WANG Yanzhen BU Hongxing NIU College of Life Sciences Henan Normal UniversityXinxiangChina School of Basic Medical Sciences Xinxiang Medical UniversityXinxiangChina

The transfer of mitochondrial DNA to the nuclear genome gives rise to the nuclear DNA sequences of mitochon-drial origin(NUMTs),considered as a driving force in genome evolution.In this study,NUMTs in 23 bat genomes were investigated and compared systematically.The results showed that NUMTs existed in 22 genomes except for Noctilio leporinus,suggesting that mitochondrial fragment insertion in the nuclear genome was a common event in bat genomes.However,remarkable variations in NUMTs number,cumulative length,and proportion in the nu-clear genome were discovered across bat species.Further orthologous NUMT loci analysis of the Phyllostomidae family indicated that the NUMTs insertion events in bat genomes were homoplasy-free.The NUMTs were mainly inserted into the intergenic regions,particularly,co-localized with repetitive sequences(especially transposable el-ements).However,several NUMTs were inserted into genes,some of which were in the exon region of functional genes.One NUMT in the genome of Pteropus alecto surprisingly matched with cDNA of ATP8B3 that provided evidence of NUMTs with coding function.Phylogenic analysis on NUMTs originating from COXI and COXII loci highlighted 2 clusters of Yinpterochiroptera and Yangochiroptera for Chiroptera.Seven NUMTs from Rhinolophus ferrumequinum were ampliﬁed,and the sequencing results conﬁrmed the reliability of the NUMT analysis.One of them was polymorphic for the presence or absence of the NUMT insertion,and each genotype of NUMT loci showed a distinct regional distribution pattern.The information obtained in this study provides novel insights into the NUMT organization and features in bat genomes and establishes a basis for further studying of the evolution of bat species.

关键词： bat homoplasy-free mitochondrial DNA NUMTs polymorphism

Global patterns of phylogenetic diversity and transmission of bat coronavirus

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Science China(Life Sciences) 2023年第4期66卷 861-874页

作者： Zhilin Wang Guangping Huang Mingpan Huang Qiang Dai Yibo Hu Jiang Zhou Fuwen Wei CAS Key Laboratory of Animal Ecology and Conservation Biology Institute of ZoologyChinese Academy of SciencesBeijing 100101China University of Chinese Academy of Sciences Beijing 100049China Chengdu Institute of Biology Chinese Academy of SciencesChengdu 610041China School of Karst Science Guizhou Normal UniversityGuiyang 550000China Center for Evolution and Conservation Biology Southern Marine Science and Engineering Guangdong Laboratory(Guangzhou)Guangzhou 511458China

bats are reservoirs for multiple coronaviruses(Co Vs).However,the phylogenetic diversity and transmission of global bat-borne Co Vs remain poorly understood.Here,we performed a Bayesian phylogeographic analysis based on 3,594 bat Co V Rd Rp gene sequences to study the phylogenetic diversity and transmission of bat-borne Co Vs and the underlying driving factors.We found that host-switching events occurred more frequently forα-Co Vs than forβ-Co Vs,and the latter was highly constrained by bat phylogeny.bat species in the families Molossidae,Rhinolophidae,Miniopteridae,and Vespertilionidae had larger contributions to the cross-species transmission of bat Co Vs.Regions of eastern and southern Africa,southern South America,Western Europe,and Southeast Asia were more frequently involved in cross-region transmission events of bat Co Vs than other regions.Phylogenetic and geographic distances were the most important factors limiting Co V transmission.bat taxa and global geographic hotspots associated with bat Co V phylogenetic diversity were identified,and bat species richness,mean annual temperature,global agricultural cropland,and human population density were strongly correlated with the phylogenetic diversity of bat Co Vs.These findings provide insight into bat Co Vevolution and ecological transmission among bat taxa.The identified hotspots of bat Co V evolution and transmission will guide early warnings of bat-borne Co V zoonotic diseases.

关键词： bat coronavirus ecological transmission phylogenetic diversity driving factors

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Lysyl oxidase inhibition enhances browning of white adipose tissue and adaptive thermogenesis

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Genes & Diseases 2022年第1期9卷 140-150页

作者： Chun Xing Duo Jiang Yang Liu Qiqun Tang Haiyan Huang Key Laboratory of Metabolism and Molecular Medicine The Ministry of EducationDepartment of Biochemistry and Molecular BiologyFudan University School of Basic Medical SciencesShanghai 200032PR China

Accumulating evidence from both animal and human studies suggests that activation of beige fat increases cellular energy expenditure,ultimately reducing adiposity.Here,we report the central role of adipocyte-derived lysyl oxidase(Lox)in the formation of thermogenic beige fat.Mice exposed to cold or aβ3 agonist showed drastically lower Lox expression in thermogenically activated beige fat.Importantly,inhibition of Lox activity with BAPN stimulated biogenesis of beige fat in inguinal white adipose tissue(iWAT)under housing conditions and potentiated cold-induced adaptive thermogenesis and beiging in both iWAT and epididymal white adipose tissue(eWAT).Notably,white adipocytes with Lox repression undergo transdifferentiation into beige adipocytes which can be suppressed by tumor necrosis factor-α(TNFα)via ERK activation.This work provides new insight into the molecular control to expand beige fat by Lox inhibition and suggest the potential for utilizing inhibitor of Lox to treat the emerging epidemics of obesity and diabetes.

关键词： Adaptive thermogenesis bat Beige fat Lysyl oxidase TNFa

EPAS1 prevents telomeric damage-induced senescence by enhancing transcription of TRF1,TRF2,and RAD50

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Zoological Research 2023年第3期44卷 636-649页

作者： Kai-Qin Li Gao-Jing Liu Xiu-Yun Liu Qiong-Fang Chen Xiao-Yan Huang Qiu Tu Jiao Zhang Qing Chang Yun-Hua Xie Rong Hua Dong-Ming Xu Zhen Liu Bo Zhao Key Laboratory of Animal Models and Human Disease Mechanisms of the Chinese Academy of Sciences&Yunnan Province KIZ-CUHK Joint Laboratory of Bioresources and Molecular Research in Common DiseasesKunming Institute of ZoologyChinese Academy of SciencesKunmingYunnan 650201China State Key Laboratory of Genetic Resources and Evolution Kunming Institute of ZoologyChinese Academy of SciencesKunmingYunnan 650201China University of Chinese Academy of Sciences Beijing 100049China Primate Facility National Research Facility for Phenotypic&Genetic Analysis of Model Animalsand National Resource Center for Non-Human PrimatesKunming Institute of ZoologyChinese Academy of SciencesKunmingYunnan 650201China

Telomeres are nucleoprotein structures located at the end of each chromosome,which function in terminal protection and genomic stability.Telomeric damage is closely related to replicative senescence in vitro and physical aging in vivo.As relatively long-lived mammals based on body size,bats display unique telomeric patterns,including the upregulation of genes involved in alternative lengthening of telomeres(ALT),DNA repair,and DNA replication.At present,however,the relevant molecular mechanisms remain unclear.In this study,we performed cross-species comparison and identified EPAS1,a well-defined oxygen response gene,as a key telomeric protector in bat fibroblasts.bat fibroblasts showed high expression of EPAS1,which enhanced the transcription of shelterin components TRF1 and TRF2,as well as DNA repair factor RAD50,conferring bat fibroblasts with resistance to senescence during long-term consecutive expansion.Based on a human single-cell transcriptome atlas,we found that EPAS1 was predominantly expressed in the human pulmonary endothelial cell subpopulation.Using in vitro-cultured human pulmonary endothelial cells,we confirmed the functional and mechanistic conservation of EPAS1 in telomeric protection between bats and humans.In addition,the EPAS1 agonist M1001 was shown to be a protective compound against bleomycin-induced pulmonary telomeric damage and senescence.In conclusion,we identified a potential mechanism for regulating telomere stability in human pulmonary diseases associated with aging,drawing insights from the longevity of bats.

关键词： bat Telomere Senescence EPAS1 M1001 Pulmonary endothelial cell

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WTAP regulates postnatal development of brown adipose tissue by stabilizing METTL3 in mice

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Life Metabolism 2022年第3期1卷 270-284页

作者： Yuqin Wang Xinzhi Li Cenxi Liu Liying Zhou Lei Shi Zhiguo Zhang Long Chen Ming Gao Lanyue Gao Yuanyuan Xu He Huang Jin Li Zheng Chen HIT Center for Life Sciences School of Life Science and TechnologyHarbin Institute of TechnologyHarbinHeilongjiang 150001China School of Life Science State Key Laboratory of Genetic EngineeringFudan UniversityShanghai 200438China Shanghai Key Laboratory of Metabolic Remodeling and Health Institute of Metabolism and Integrative BiologyFudan UniversityShanghai 200438China Department of Cardiology The First Hospital of Jilin UniversityChangchunJilin 130021China The Key Laboratory of Liaoning Province on Toxic and Biological Effects of Arsenic School of Public HealthChina Medical UniversityShenyangLiaoning 110122China

Brown adipocyte maturation during postnatal development is essential for brown adipose tissue(bat)to protect animals against cold.Impaired maturation of brown adipocytes leads to cold intolerance.However,the molecular mechanisms that determine the maturation of brown adipocytes during postnatal development are not fully understood.Here,we identify Wilms’tumor 1-associating protein(WTAP)as an essential regulator in the postnatal development and maturation of bat.bat-specific knockout of Wtap(Wtap-BKO)severely impairs maturation of bat in vivo by decreasing the expression of bat-selective genes,leading to the whitening of interscapular bat(ibat).Single nucleus RNA-sequencing analysis shows the dynamic changes of cell heterogeneity in ibat of Wtap-BKO mice.Adult mice with WTAP deficiency in bat display hypothermic and succumb to acute cold challenge.Mechanistically,WTAP deficiency decreases m6A mRNA modification by reducing the protein stability of METTL3.bat-specific overexpression of Mettl3 partially rescues the phenotypes observed in Wtap-BKO mice.These data demonstrate that WTAP/METTL3 plays an essential role in ibat postnatal development and thermogenesis.

关键词： WTAP bat m6A postnatal development METTL3 PRDM16

Site-specific conjugation of bifunctional chelator bat to mouse IgG1 Fab~1 fragment

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Acta Pharmacologica Sinica 2006年第2期27卷 237-241页

作者： Jun LI~1 Xue-hao WANG Xiao-ming WANG Zhao-lai CHEN Division of Hepatic Surgery.Department of Surgery,the First Affiliated Hospital of Nanjing Medical University and Jiangsu Province Hospital.Nanjing 210029.China Division of Hepatic Surgery Department of Surgery the First Affiliated Hospital of Nanjing Medical University and Jiangsu Province Hospital Nanjing China

Aim:To perform a site-specific conjugation of Fab’ fragments of a mouse mono-clonal antibody(MoAb)B43(of IgG1subtype)to a bifunctional chelator 6-[p-(bromoacetamido)benzyl]-1,4,8,11-tetraazacyclotetradecane-N,N′,N″,N′″-tetraacetic acid(bat)via the thiol groups in the hinge distal to the antigen-binding site of the Fab~1.Methods:B43 was cleaved using a simple 2-step method.First,stable F(ab~1)2was produced by pepsin treatment.Fab~1 with tree thiol in thehinge region was then obtained by cysteine reduction of F(ab~1)2.Second,a site-specific conjugation of Fab~1 to thiol-specific bat was performed in a one-stepreaction.Results:The Fab~1 fragment had approximately 1.8 free thiol groups permolecule after cysteine reduction.The conjugation efficiency and the chemicalyield were approximately 1.28 moles chelator/Fab′ and 74% of the initial concentra-tion of Fab′,respectively.The F(ab′)2,Fab′ and Fab′-bat all maintained reason-able antigen-binding properties.67Cu labeling of the conjugate under standardconditions did not impair the immunoreactivity of Fab′-bat.Conclusion:This isa simple and efficient method for producing immunoreactive conjugates of Fab′-bat,which can be used to make radiometal-labeled conjugates for further diag-nostic and therapeutic applications.

关键词： conjugation bat chelator monoclonal antibody

An overview of energy and metabolic regulation

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Science China(Life Sciences) 2019年第6期62卷 771-790页

作者： Song Wen Chaoxun Wang Min Gong Ligang Zhou Department of Endocrinology Shanghai Pudong Hospital Fudan University

The physiology and behaviors related to energy balance are monitored by the nervous and humoral systems. Because of the difficulty in treating diabetes and obesity, elucidating the energy balance mechanism and identifying critical targets for treatment are important research goals. Therefore, the purpose of this article is to describe energy regulation by the central nervous system(CNS) and peripheral humoral pathway. Homeostasis and rewarding are the basis of CNS regulation. Anorexigenic or orexigenic effects reflect the activities of the POMC/CART or NPY/AgRP neurons within the hypothalamus. Neurotransmitters have roles in food intake, and responsive brain nuclei have different functions related to food intake, glucose monitoring, reward processing. Peripheral gut-or adipose-derived hormones are the major source of peripheral humoral regulation systems. Nutrients or metabolites and gut microbiota affect metabolism via a discrete pathway. We also review the role of peripheral organs, the liver,adipose tissue, and skeletal muscle in peripheral regulation. We discuss these topics and how the body regulates metabolism.

关键词： hedonic POMC/CART NPY/AgRP neurotransmitter brain nuclei gut-derived hormone gut microbiota bat

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