Crosslinking of substrates occurs exclusively to the p66 subunit of heterodimeric HIV-1 reverse transcriptase.
Photoaffinity labeling of the hetero- and homodimeric forms of HIV-1 reverse transcriptase has been carried out using [32P]rA12-18.dT10 as a representative template-primer and [alpha-32P]dTTP as a representative 2'-deoxynucleoside-5'-triphosphate. UV irradiation produces stable, covalent crosslinks between each of the reactants and both the hetero-(p66/p51) and homodimeric (p66/p66, p51/p51) forms of the enzyme. In the case of the p66/p51 heterodimer, the form of the enzyme believed to be involved in viral replication, crosslinking occurs exclusively to the p66 subunit. These results suggest that the polymerase activity of the heterodimer residues on p66.
Cheng
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Division of Virology, Burroughs Wellcome Co., Research Triangle Park, NC 27709.
Painter
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Furman
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D000345:Affinity Labels / Q000378:metabolism; D003001:Cloning, Molecular; D003432:Cross-Linking Reagents; D004926:Escherichia coli / Q000235:genetics; D015497:HIV-1 / Q000201:enzymology* / Q000502:physiology; D007700:Kinetics; D046911:Macromolecular Substances; D008970:Molecular Weight; D011485:Protein Binding; D012194:RNA-Directed DNA Polymerase / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D013942:Thymine Nucleotides / Q000378:metabolism; D014779:Virus Replication
D000345:Affinity Labels; D003432:Cross-Linking Reagents; D046911:Macromolecular Substances; D011994:Recombinant Proteins; D013942:Thymine Nucleotides; D012194:RNA-Directed DNA Polymerase; C024157:thymidine 5'-triphosphate