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Construction and Characterization of an Infectious Molecular Clone of Koala Retrovirus

Takayuki Shojima,a Shigeki Hoshino,a Masumi Abe,b Jiro Yasuda,b,c Hiroko Shogen,a Takeshi Kobayashi,a Takayuki Miyazawaa

Laboratory of Signal Transduction, Department of Cell Biology, Institute for Virus Research, Kyoto University, Kyoto, Japana; Fifth Biology Section for Microbiology, First Department of Forensic Science, National Research Institute of Police Science, Kashiwa City, Chiba, Japanb; Department of Emerging Infectious Diseases, Institute of Tropical Medicine, Nagasaki University, Nagasaki, Japanc

ABSTRACT
Koala retrovirus (KoRV) is a gammaretrovirus that is currently endogenizing into koalas. Studies on KoRV infection have been hampered by the lack of a replication-competent molecular clone. In this study, we constructed an infectious molecular clone, termed plasmid pKoRV522, of a KoRV isolate (strain Aki) from a koala reared in a Japanese zoo. The virus KoRV522, derived from pKoRV522, grew efficiently in human embryonic kidney (HEK293T) cells, attaining 106 focus-forming units/ml. Several mutations in the Gag (L domain) and Env regions reported to be involved in reduction in viral infection/production in vitro are found in pKoRV522, yet KoRV522 replicated well, suggesting that any effects of these mutations are limited. Indeed, a reporter virus pseudotyped with pKoRV522 Env was found to infect human, feline, and mink cell lines efficiently. Analyses of KoRV Ldomain mutants showed that an additional PPXY sequence, PPPY, in Gag plays a critical role in KoRV budding. Altogether, our results demonstrate the construction and characterization of the first infectious molecular clone of KoRV. The infectious clone reported here will be useful for elucidating the mechanism of endogenization of the virus in koalas and screening for antiretroviral drugs for KoRV-infected koalas.

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  • 2013
  • Biogeography
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  • Chlamydia
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  • Ellis
  • Eucalyptus
  • Genetics
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