Molecular Characterization of HIV-2 (ROD) Protease Following PCR Cloning from Virus Infected H9 Cells
A 450 nucleotide sequence corresponding to the nucleotides 1931-2380 of the viral genome (8) was amplified by polymerase chain reaction (PCR) using template DNA prepared from HIV-2 (ROD) infected H9 cells. The sequence codes for HIV-2 protease and its N-terminal flanking peptide. An identical DNA sequence was obtained from three independent PCR amplifications, which differs from the published sequence of HIV-2 (ROD) in 7 nucleotides scattered throughout the region of the cloned DNA. The cloned DNA was expressed in E. coli cells and resulted in the synthesis of a correctly processed HIV-2 protease, which is enzymatically active. Therefore, none of the seven nucleotide changes, which resulted in two amino acid substitutions, affect the autoproteolytic or trans-cleaving activities of the HIV-2 protease.
KeywordsHuman Immunodeficiency Virus Human Immunodeficiency Virus Type Amino Acid Change Human Immunodeficiency Virus Protease Human Immunodeficiency Virus Proteinase Inhibitor
Unable to display preview. Download preview PDF.
- Le Grice, S.F.J., Roswitha, E., Mills, J., and Mous, J. 1989. Comparison of the human immunodeficiency virus type 1 and 2 proteases by hybrid gene construction and trans-complementation. J. Biol. Chem. 364:14902–8.Google Scholar
- Pichuantest, S., Babe, L.M., Barr, P.J., DeCamp, D.L., and Craik, C.S. 1990. Recombinant HIV2 protease processes HIV1 Pr53 gag and analogous junction peptides in vitro. J. Biol. Chem. 265:13890–8.Google Scholar
- Rittenhouse, J., Turon, M.C., Helfrich, R.J., Albrecht, K.S., Weigl, D., Simmer, R.L., Mordini, F., Erickson, J., and Kohlbrenner, W.E. 1990. Affinity purification of HIV-1 and HIV-2 proteases from recombinant E. Coli strains using pepstatinagarose. Biochem. Biophys. Acad. Comm. 171:60–6.CrossRefGoogle Scholar
- Roberts, N.A., Martin, J.A., Kinchington, D., Broadhurst, A.V., Craig, J.C., Duncan, I.B., Galpin, S.A., Handa, B.K., Kay, J., Krohn, A., Lambert, R.W., Merrett, J.H., Mills, J.S., Parkes, K.E.B., Redshaw, S., Ritchie, A.J., Taylor, D.L., Thomas, G.J., Machin, P.J. 1990. Rational design of peptide-based HIV proteinase inhibitors. Science 248:358–61.PubMedCrossRefGoogle Scholar