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Research Article

A tick-borne segmented RNA virus contains genome segments derived from unsegmented viral ancestors

Xin-Cheng Qin, Mang Shi, Jun-Hua Tian, Xian-Dan Lin, Dong-Ya Gao, Jin-Rong He, Jian-Bo Wang, Ci-Xiu Li, Yan-Jun Kang, Bin Yu, Dun-Jin Zhou, Jianguo Xu, Alexander Plyusnin, Edward C. Holmes, and Yong-Zhen Zhang
PNAS May 6, 2014 111 (18) 6744-6749; first published April 21, 2014; https://doi.org/10.1073/pnas.1324194111
Xin-Cheng Qin
aState Key Laboratory for Infectious Disease Prevention and Control, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Department of Zoonoses, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100206, China;
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Mang Shi
aState Key Laboratory for Infectious Disease Prevention and Control, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Department of Zoonoses, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100206, China;
bMarie Bashir Institute for Infectious Diseases and Biosecurity, Charles Perkins Centre, School of Biological Sciences and Sydney Medical School, University of Sydney, Sydney, NSW 2006, Australia;
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Jun-Hua Tian
cWuhan Center for Disease Control and Prevention, Wuhan 430015, China;
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Xian-Dan Lin
dWenzhou Center for Disease Control and Prevention, Wenzhou 325001, China; and
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Dong-Ya Gao
aState Key Laboratory for Infectious Disease Prevention and Control, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Department of Zoonoses, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100206, China;
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Jin-Rong He
aState Key Laboratory for Infectious Disease Prevention and Control, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Department of Zoonoses, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100206, China;
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Jian-Bo Wang
aState Key Laboratory for Infectious Disease Prevention and Control, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Department of Zoonoses, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100206, China;
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Ci-Xiu Li
aState Key Laboratory for Infectious Disease Prevention and Control, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Department of Zoonoses, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100206, China;
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Yan-Jun Kang
aState Key Laboratory for Infectious Disease Prevention and Control, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Department of Zoonoses, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100206, China;
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Bin Yu
cWuhan Center for Disease Control and Prevention, Wuhan 430015, China;
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Dun-Jin Zhou
cWuhan Center for Disease Control and Prevention, Wuhan 430015, China;
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Jianguo Xu
aState Key Laboratory for Infectious Disease Prevention and Control, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Department of Zoonoses, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100206, China;
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Alexander Plyusnin
aState Key Laboratory for Infectious Disease Prevention and Control, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Department of Zoonoses, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100206, China;
eDepartment of Virology, Haartman Institute, University of Helsinki, 00014, Helsinki, Finland
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Edward C. Holmes
aState Key Laboratory for Infectious Disease Prevention and Control, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Department of Zoonoses, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100206, China;
bMarie Bashir Institute for Infectious Diseases and Biosecurity, Charles Perkins Centre, School of Biological Sciences and Sydney Medical School, University of Sydney, Sydney, NSW 2006, Australia;
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Yong-Zhen Zhang
aState Key Laboratory for Infectious Disease Prevention and Control, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Department of Zoonoses, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100206, China;
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  • For correspondence: zhangyongzhen@icdc.cn
  1. Edited by Peter Palese, Icahn School of Medicine at Mount Sinai, New York, NY, and approved March 27, 2014 (received for review December 30, 2013)

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    Fig. 1.

    Transmission EM image of JMTV in (A) infected cells and (B) cell culture supernatant. The viral particles are indicated by the orange arrowheads.

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    Fig. 2.

    Schematic genome organization and putative proteomic maps of JMTV, showing predicted ORFs (blue boxes), transmembrane regions (yellow boxes), signal peptides (red box), and N-linked glycosylation sites (upward-pointing pink triangles).

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    Fig. 3.

    ML phylogenetic trees of (A) NS3 and (B) NS5 proteins of JMTV and TCLA within the context of representative flaviviruses. The trees are midpoint-rooted, and all horizontal branches are drawn to a scale of amino acid substitutions per site. The branch and taxon labels of the JMTV–TCLA clade are shaded red. For clarity, bootstrap values are only shown for internal nodes.

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    Fig. 4.

    ML phylogenetic trees of the four genome segments obtained from nine depooled JMTV samples. The trees are midpoint-rooted, and all horizontal branches are drawn to a scale of nucleotide substitutions per site. Major clade/lineages are shaded with blue, yellow, and red, respectively. Phylogenetic incongruence, suggestive of reassortment, is indicated with colored arrowheads.

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    Table 1.

    Homology search results and expression levels of JMTV genome segments in the context of total RNA

    ContigTop BLAST and HHblits hitsLength, nt*No. of mapped readsExpression level**Total reads, %Non-rRNA reads, %
    comp194ANT5, NS5 (TABV)3,072980,35915,956.364.4418.67
    comp196—2,785961,71917,266.054.3618.32
    comp197ANT34, NS3 (TABV)2,793767,02013,731.113.4814.61
    comp202ANT3.12,751459,8748,358.312.088.76
    Total11,4013,168,97251,696.3414.3760.36
    • ↵* Length (measured in nt) of genomic RNAs determined after cyclization RT-PCR, excluding poly(A) tail.

    • † Expression level is calculated by using length of mapped reads/length of contig.

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    Table 2.

    Prevalence of JMTV in ticks, mosquitoes, and cattle

    SpeciesLocation*Mammalian hostNo. of pools testedNo. of positive pools
    R. microplus1–7Cattle181114
    H. longicornis1Dog, goat, cattle9150
    H. campanulata8Dog2418
    H. flava7–9Hedgehog, badger455
    I. sinesis1Wild goat11
    I. granulatus10NA31
    R. sanguineus11NA31
    Haemaphysalis hystricis9Badger320
    Armigeres sp. (mosquito)1NK201
    Culex sp. (mosquito)1NK310
    Anopheles sp. (mosquito)1NK50
    Bos sp. (cattle)1, 6, 10NK1455
    Total——481196
    • NA, not applicable; NK, not known.

    • ↵* Locations: 1, Wuhan; 2, Shiyan; 3, Shengnongjia; 4, Yichang; 5, Enshi; 6, Jingmen; 7, Xiangfan; 8, Xinzhou; 9, Huanggang; 10, Wenzhou; 11, Ningbo.

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Segmented relative of flavivirus
Xin-Cheng Qin, Mang Shi, Jun-Hua Tian, Xian-Dan Lin, Dong-Ya Gao, Jin-Rong He, Jian-Bo Wang, Ci-Xiu Li, Yan-Jun Kang, Bin Yu, Dun-Jin Zhou, Jianguo Xu, Alexander Plyusnin, Edward C. Holmes, Yong-Zhen Zhang
Proceedings of the National Academy of Sciences May 2014, 111 (18) 6744-6749; DOI: 10.1073/pnas.1324194111

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Segmented relative of flavivirus
Xin-Cheng Qin, Mang Shi, Jun-Hua Tian, Xian-Dan Lin, Dong-Ya Gao, Jin-Rong He, Jian-Bo Wang, Ci-Xiu Li, Yan-Jun Kang, Bin Yu, Dun-Jin Zhou, Jianguo Xu, Alexander Plyusnin, Edward C. Holmes, Yong-Zhen Zhang
Proceedings of the National Academy of Sciences May 2014, 111 (18) 6744-6749; DOI: 10.1073/pnas.1324194111
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