Expression of the heterodimeric form of human immunodeficiency virus type 2 reverse transcriptase in Escherichia coli and characterization of the enzyme.
A system for the expression of recombinant human immunodeficiency virus type 2 (HIV-2) reverse transcriptase (RT) in Escherichia coli has been developed, which allows purification of the heterodimeric form of the enzyme as well as the separate purification of the two subunits. It is shown that equilibrium formation between monomeric and homodimeric forms of the recombinant 66- and 51-kDa subunits is considerably more rapid than in the case of the corresponding homodimeric forms of HIV-1 RT. In accordance with our previously published studies on HIV-1 RT (Restle, T., Müller, B., and Goody, R.S. (1990) J. Biol. Chem. 265, 8986-8988) RNA-dependent DNA polymerase activity of the HIV-2 RT preparations can be exactly correlated to their dimer content. No significant heterodimer formation can be observed upon coexpression of the 66-kDa subunit of HIV-2 RT with the 51-kDa subunit of HIV-1 RT in the same cell, indicating differences in the dimerization domains of the two proteins. Recombinant HIV-2 RT is not recognized by a set of 23 monoclonal antibodies raised against HIV-1 RT, although it shows weak cross-reactivity with sera from HIV-1-infected patients.
Müller
B
B
Abteilung Biophysik, Max Planck Institut für medizinische Forschung, Heidelberg, Federal Republic of Germany.
Restle
T
T
Kühnel
H
H
Goody
R S
RS
D002850:Chromatography, Gel; D003001:Cloning, Molecular; D003429:Cross Reactions; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D005798:Genes, Bacterial; D005822:Genetic Vectors; D015497:HIV-1 / Q000201:enzymology; D015498:HIV-2 / Q000201:enzymology*; D006801:Humans; D007700:Kinetics; D012194:RNA-Directed DNA Polymerase / Q000235:genetics* / Q000378:metabolism
D012194:RNA-Directed DNA Polymerase