Generation and characterization of a stable full-length ecotropic murine leukemia virus molecular clone that produces novel phenotypes to Fv1 restriction.
Retrovirus tropism can be restricted by host cell factors such as Fv1, TRIM5alpha, and Lv1 that inhibit infection by targeting the incoming viral capsid. The Fv1 gene inhibits murine leukemia virus infection in mice, but the precise mechanism of Fv1-mediated restriction is poorly understood. Our previous studies had demonstrated that Fv1-mediated viral tropism can be determined within the capsid protein at position 114 (Jung and Kozak. 2000. J. Virol. 74: 5385-7). To study the interaction between Fv1 and CA, we introduced amino acid substitution and deletion at this site in the N-tropic AKV capsid gene. The mutated two-LTR proviral DNAs were introduced into SC-1 cells by transfection. After transfection, cell supernatants collected from transfected cells were tested for host range susceptibility. The result indicated that substitution of amino acids did not alter tropism, but the deletion of 114His produced a virus with unusual tropism. The novel phenotype produced here failed to replicate in Fv1-expressing cells. This mutant virus showing such an extreme restriction pattern would be useful for studying the mechanism of Fv1- mediated restriction.
Bae
Eun Hye
EH
Department of Microbiology and Institute of Basic Science, Dankook University, Cheonan 330-714, Korea.
Park
Sung-Han
SH
Park
Sang Min
SM
Park
Jin Woo
JW
Lim
Mi Suk
MS
Jung
Yong-Tae
YT
D019943:Amino Acid Substitution; D000818:Animals; D036022:Capsid Proteins / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D015683:Gene Products, gag / Q000235:genetics / Q000378:metabolism*; D009052:Leukemia Virus, Murine / Q000235:genetics / Q000502:physiology*; D051379:Mice; D008807:Mice, Inbred BALB C; D016297:Mutagenesis, Site-Directed; D041681:NIH 3T3 Cells; D011506:Proteins / Q000235:genetics / Q000378:metabolism*; D020079:Terminal Repeat Sequences; D014162:Transfection; D018523:Tropism; D014779:Virus Replication
D036022:Capsid Proteins; C103189:Fv1 protein, mouse; D015683:Gene Products, gag; D011506:Proteins