| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
1 National Centre for HIV Virology Research, Institute of Medical and Veterinary Science, Frome Road, Adelaide 5000, South Australia
and2 Department of Microbiology and Immunology, University of Adelaide, North Terrace, Adelaide 5000, South Australia, Australia
We have investigated the kinetics of human immuno-deficiency virus (HIV) reverse transcription in infected T cells, using a synchronized, one-step, cell-to-cell infection model and quantitative PCR assays for the different DNA intermediate structures that are found sequentially during reverse transcription. Different efficiencies that might arise from the use of different primers and other PCR conditions were normalized by conversion of each PCR product signal to copy numbers by comparing with standards. After an initial lag period, the minus-strand strong-stop viral DNA was detected first. This was followed by the post-transfer newly extended minus-strand viral DNA and then by the plus-strand strong-stop DNA and fully extended minus-strand DNA. Kinetic data indicated that, once reverse transcription was initiated, the HIV reverse transcriptase synthesized minus-strand DNA at a rate of 150–180 bases/min, and that the first template transfer and the initiation of the plus-strand DNA synthesis imposed specific time delays. In contrast, minus-strand viral DNA synthesized after the second template transfer appeared at a time point very close to the time of the appearance of the last piece of DNA synthesized just before the second template switch, suggesting that the second switch occurred very rapidly. Taken together, our results define more accurately than was previously possible the rates of several of the steps in HIV reverse transcription in infected T cell lines and indicate different mechanisms for the two distinct template switches during retrovirus reverse transcription.
* Author for correspondence. Fax +61 8 228 7538. e-mail jmcinnes@microb.adelaide.edu.au
Received 14 November 1994;
accepted 27 February 1995.
This article has been cited by other articles:
|
|
 |
|
  D. C. Thomas, Y. A. Voronin, G. N. Nikolenko, J. Chen, W.-S. Hu, and V. K. Pathak Determination of the Ex Vivo Rates of Human Immunodeficiency Virus Type 1 Reverse Transcription by Using Novel Strand-Specific Amplification Analysis J. Virol., May 1, 2007; 81(9): 4798 - 4807. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Sourisseau, N. Sol-Foulon, F. Porrot, F. Blanchet, and O. Schwartz Inefficient Human Immunodeficiency Virus Replication in Mobile Lymphocytes J. Virol., January 15, 2007; 81(2): 1000 - 1012. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Jolly and Q. J. Sattentau Human Immunodeficiency Virus Type 1 Virological Synapse Formation in T Cells Requires Lipid Raft Integrity J. Virol., September 15, 2005; 79(18): 12088 - 12094. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. Vandegraaff, R. Kumar, C. J. Burrell, and P. Li Kinetics of Human Immunodeficiency Virus Type 1 (HIV) DNA Integration in Acutely Infected Cells as Determined Using a Novel Assay for Detection of Integrated HIV DNA J. Virol., November 15, 2001; 75(22): 11253 - 11260. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Auxilien, G. Keith, S. F. J. Le Grice, and J.-L. Darlix Role of Post-transcriptional Modifications of Primer tRNALys,3 in the Fidelity and Efficacy of Plus Strand DNA Transfer during HIV-1 Reverse Transcription J. Biol. Chem., February 12, 1999; 274(7): 4412 - 4420. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
| INT J SYST EVOL MICROBIOL | MICROBIOLOGY | J GEN VIROL |
| J MED MICROBIOL | ALL SGM JOURNALS | |
