Expression and frameshifting but extremely inefficient proteolytic processing of the HIV-1 gag and pol gene products in stably transfected rodent cell lines.
Expression, ribosomal frameshifting, and proteolytic processing of HIV-1 GAG and POL proteins were investigated in heterologous mammalian cells in order to elucidate the influence of the cellular background on these events. DNA fragments encoded by the gag and pol region were expressed in two rodent cell lines, LTK- and BHK. Both stably transfected cell lines continuously produce recombinant proteins which react with HIV-specific antisera. The GAG precursor and a 39-kDa proteolytic fragment thereof were the major recombinant proteins detected. Expression of the gag-pol region leads to the production of the GAG-POL precursor. Ribosomal frameshifting at the HIV-1 shifty sequence to a typical extent could be positively demonstrated by an enzyme assay. Despite the presence of the viral protease within the GAG-POL precursors, proteolytic processing of the HIV-derived polyproteins was extremely inefficient. The efficiency could not be enhanced by overexpression of the HIV-1 protease encoding region.
Moosmayer
D
D
Deutsches Primatenzentrum, Abt. Virologie und Immunologie, Göttingen.
Reil
H
H
Ausmeier
M
M
Scharf
J G
JG
Hauser
H
H
Jentsch
K D
KD
Hunsmann
G
G
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002522:Chlorocebus aethiops; D006224:Cricetinae; D016368:Frameshift Mutation; D015870:Gene Expression; D015683:Gene Products, gag / Q000235:genetics* / Q000378:metabolism; D015684:Gene Products, pol / Q000235:genetics* / Q000378:metabolism; D016333:HIV Protease / Q000378:metabolism; D015497:HIV-1 / Q000235:genetics*; D006801:Humans; D006868:Hydrolysis; D051379:Mice; D008969:Molecular Sequence Data; D011499:Protein Processing, Post-Translational; D013379:Substrate Specificity; D014162:Transfection
D015683:Gene Products, gag; D015684:Gene Products, pol; D016333:HIV Protease