Transdominant inhibition of Moloney murine leukemia virus proliferation by defective mutants of reverse transcriptase.
To investigate the dominant-negative inhibition of Moloney murine leukemia virus (Mo-MuLV) proliferation by polymerization-defective mutants of reverse transcriptase (RT), we constructed several plasmids harboring the Mo-MuLV RT gene in which the YVDD sequence, one of the conserved sequences in RNA-dependent polymerases, was altered, and then transformed mouse NIH3T3 cells and Escherichia coli with the mutant plasmids. Mouse NIH3T3 cells expressing these mutant RT genes were highly resistant to Mo-MuLV proliferation. The mutant RT expressed in E. coli exhibited no polymerization activity, but it retained its binding activity to the template RNA and inhibited in vitro poly(dG) synthesis occurring with the wild-type RT. These results suggest that the competition for binding of the two types of enzymes to the template is responsible for the resistance to Mo-MuLV proliferation and that the YVDD sequence of Mo-MuLV may be a good target for dominant-negative inhibition of retroviral proliferation.
Shirasawa
Y
Y
Yakult Central Institute for Microbiological Research, Kunitachi, Tokyo.
Sato
S
S
Osawa
T
T
Hirashima
A
A
D016475:3T3 Cells / Q000378:metabolism / Q000821:virology; D000595:Amino Acid Sequence; D000818:Animals; D017124:Conserved Sequence; D018076:DNA, Complementary / Q000037:antagonists & inhibitors / Q000096:biosynthesis; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005814:Genes, Viral / Q000235:genetics; D051379:Mice; D008979:Moloney murine leukemia virus / Q000201:enzymology* / Q000235:genetics / Q000502:physiology; D009154:Mutation; D010957:Plasmids / Q000235:genetics; D011061:Poly A / Q000378:metabolism; D011068:Poly G / Q000037:antagonists & inhibitors / Q000096:biosynthesis; D011485:Protein Binding; D012194:RNA-Directed DNA Polymerase / Q000235:genetics / Q000378:metabolism*; D014162:Transfection; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism; D014779:Virus Replication
D018076:DNA, Complementary; D014764:Viral Proteins; D011061:Poly A; D011068:Poly G; C026628:poly(dG); D012194:RNA-Directed DNA Polymerase