Affinity modification of human immunodeficiency virus reverse transcriptase and DNA template by photoreactive dCTP analogs

FEBS Lett. 1994 Nov 7;354(2):200-2. doi: 10.1016/0014-5793(94)01110-9.


New base-substituted analogs of dCTP containing an azido group have been synthesized and applied to a selective photoaffinity modification of HIV-RT (p66/p51 heterodimer). The labeling of only the 66 kDa subunit of HIV-RT was detected when the enzyme was first irradiated with the analogs and then template (5'-(d)GGTTAAATAAAATAGTAAGAATGTATAGCCCCTACCA-3') and 5' 32P end-labeled 3'-(d)TTACATATCGGGGATGGT-5' primer were added. The 5' 32P end-labeled primer elongated by dCTP analogs in the presence of both HIV-RT and DNA template is able to modify both subunits of HIV-RT and DNA template. This way of specific cross-linking to both DNA (RNA) template and HIV-RT opens up new possibilities to study the HIV-RT active site.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Affinity Labels*
  • Base Sequence
  • Binding Sites
  • DNA / chemistry*
  • DNA / metabolism
  • Deoxycytosine Nucleotides / chemistry
  • Deoxycytosine Nucleotides / metabolism
  • Deoxycytosine Nucleotides / pharmacology*
  • HIV / enzymology*
  • Molecular Sequence Data
  • Molecular Structure
  • Photochemistry
  • RNA-Directed DNA Polymerase / chemistry*
  • RNA-Directed DNA Polymerase / metabolism
  • Templates, Genetic
  • Ultraviolet Rays


  • 5-(3-(4-azidotetrafluorobenzamido)allyl)-2'-deoxycytidine-5'-triphosphate
  • Affinity Labels
  • Deoxycytosine Nucleotides
  • exo-N-(2-(4-azidotetrafluorobenzamido)ethyl)-deoxycytidine-5'-triphosphate
  • 2'-deoxycytidine 5'-triphosphate
  • DNA
  • RNA-Directed DNA Polymerase