The DNA replication, virogenesis and infection of canine minute virus in non-permissive and permissive cells.
Canine minute virus (CnMV), a kind of autonomous parvovirus, is a member of genus bocavirus in parvovirdae family. In our previous study, we constructed and obtained infectious clones of CnMV, analyzed genome characteristics, RNA transcription profile, and revealed some molecular mechanisms of cytopathic effect of target cells. The purpose of this study was to investigate DNA replication, virogenesis and infectious tropism of CnMV in non-permissive and permissive cells. We demonstrated that the genomic DNA of CnMV, besides WRD cells, could replicate significantly in some non-permissive cells (CrFK, EBtR and COS-7) following transfection with infectious clone of CnMV, pI-MVC. Moreover, by using Western blotting and immunofluorescence, we found that the NS1 protein of CnMV was obviously expressed in both 293, CrFK, EBtR and COS-7 cells transfected with pI-MVC. Meanwhile, two-rounds of reinfection on WRD cells (blind passage) of the transfected cell lysates in CrFK, EBtR and COS-7 cells tranfected with pI-MVC showed that pI-MVC could produce infectious virions in these types of non-permissive cells. Furthermore, it is confirmed that CnMV only infected WRD cells (permissive cells for CnMV), could not infect any non-permissive cells including CrFK, EBtR, COS-7, HK293, A549 and A9 cells. Taken together, for the first time, we have demonstrated that bocavirus CnMV DNA could replicate and form infectious progeny virus in some non-permissive cells. And what is more, unlike other parvoviruses, CnMV did not infect some non-permissive cells, although the DNA replication of CnMV occurred in these cells.
Affiliated Hospital of Ningxia Medical University, Yinchuan, Ningxia, China.
Fang
F
Li
Department of Biochemistry and Molecular Biology, Ningxia Medical University, Yinchuan, Ningxia, China.
Qian
Q
Zhang
Department of Biochemistry and Molecular Biology, Ningxia Medical University, Yinchuan, Ningxia, China.
Qing
Q
Yao
Department of Biochemistry and Molecular Biology, Ningxia Medical University, Yinchuan, Ningxia, China.
Long
L
Chen
Department of Biochemistry and Molecular Biology, Ningxia Medical University, Yinchuan, Ningxia, China.
Jianning
J
Li
Department of Microbiology, Molecular Genetics and Immunology, University of Kansas Medical Center, Kansas City, KS 66160 USA. Electronic address: jqiu@kumc.edu.
Jianming
J
Qiu
Department of Biochemistry and Molecular Biology, Ningxia Medical University, Yinchuan, Ningxia, China; Key Lab of Reproduction and Heredity of Ningxia Hui Autonomous Region, Ningxia Medical University, Yinchuan, Ningxia, China. Electronic address: sunraining2008@hotmail.com.
Yuning
Y
Sun
D000818:Animals; D052660:Bocavirus / Q000235:genetics / Q000254:growth & development / Q000502:physiology*; D002460:Cell Line; D002522:Chlorocebus aethiops; D004261:DNA Replication; D004283:Dog Diseases / Q000821:virology*; D004285:Dogs; D006801:Humans; D010322:Parvoviridae Infections / Q000662:veterinary* / Q000821:virology*; D014779:Virus Replication
Bocavirus; Canine minute virus; DNA replication; Infection; Non-permissive cells; Permissive cells; Virogenesis