Differences in Reverse Transcriptase Activity versus p24 Antigen Detection in Cell Culture, When Comparing a Homogeneous Group of HIV Type 1 Subtype B Viruses with a Heterogeneous Group of Divergent Strains

1998 ◽  
Vol 14 (4) ◽  
pp. 347-352 ◽  
Author(s):  
GARY E. CORRIGAN ◽  
LUBNA AL-KHALILI ◽  
ANDERS MALMSTEN ◽  
RIGMOR THORSTENSSON ◽  
EVA MARIA FENYÖ ◽  
...  
Keyword(s):  
Cell Culture ◽  
Reverse Transcriptase ◽  
Reverse Transcriptase Activity ◽  
Heterogeneous Group ◽  
Homogeneous Group ◽  
Transcriptase Activity ◽  
Subtype B ◽  
P24 Antigen ◽  
Hiv Type 1

Rapid Screening of Phenotypic Resistance to Nevirapine by Direct Analysis of HIV Type 1 Reverse Transcriptase Activity in Plasma

1999 ◽  
Vol 15 (13) ◽  
pp. 1191-1200 ◽  
Author(s):  
Guillermo Vazquez-Rosales ◽  
J. Gerardo Garcia Lerma ◽  
Shinji Yamamoto ◽  
William M. Switzer ◽  
Diane Havlir ◽  
...  
Keyword(s):  
Reverse Transcriptase ◽  
Direct Analysis ◽  
Reverse Transcriptase Activity ◽  
Rapid Screening ◽  
Phenotypic Resistance ◽  
Transcriptase Activity ◽  
Hiv Type 1

scholarly journals Reversion of a Human Immunodeficiency Virus Type 1 Matrix Mutation Affecting Gag Membrane Binding, Endogenous Reverse Transcriptase Activity, and Virus Infectivity

1999 ◽  
Vol 73 (6) ◽  
pp. 4728-4737 ◽  
Author(s):  
Rosemary E. Kiernan ◽  
Akira Ono ◽  
Eric O. Freed
Keyword(s):  
Human Immunodeficiency Virus ◽  
Amino Acid ◽  
Reverse Transcriptase ◽  
Reverse Transcriptase Activity ◽  
Wild Type ◽  
Viral Dna ◽  
Transcriptase Activity ◽  
Immunodeficiency Virus ◽  
Infected Cells

ABSTRACT We previously characterized mutations in the human immunodeficiency virus type 1 matrix (MA) protein that displayed reduced infectivity in single-round assays, defects in the stable synthesis of viral DNA in infected cells, and impaired endogenous reverse transcriptase activity. The mutants, which contained substitutions in a highly conserved Leu at MA amino acid 20, also increased binding of Gag to membrane. To elucidate further the role of MA in the virus replication cycle, we have characterized a viral revertant of an amino acid 20 mutant (20LK). The revertant virus, which replicates with essentially wild-type kinetics in H9 cells, contains second-site compensatory changes at MA amino acids 73 (E→K) and 82 (A→T), while retaining the original 20LK mutation. Single-cycle infectivity assays, performed with luciferase-expressing viruses, show that the 20LK/73EK/82AT triple mutant displays markedly improved infectivity relative to the original 20LK mutant. The stable synthesis of viral DNA in infected cells is also significantly increased compared with that of 20LK DNA. Furthermore, activity of revertant virions in endogenous reverse transcriptase assays is restored to near-wild-type-levels. Interestingly, although 20LK/73EK/82AT reverses the defects in replication kinetics, postentry events, and endogenous reverse transcriptase activity induced by the 20LK mutation, the reversion does not affect the 20LK-imposed increase in Gag membrane binding. Mutants containing single and double amino acid substitutions were constructed, and their growth kinetics were examined. Only virus containing all three changes (20LK/73EK/82AT) grew with significantly accelerated kinetics; 73EK, 73EK/82AT, and 20LK/82AT mutants displayed pronounced defects in virus particle production. Viral core-like complexes were isolated by sucrose density gradient centrifugation of detergent-treated virions. Intriguingly, the protein composition of wild-type and mutant detergent-resistant complexes differed markedly. In wild-type and 20LK complexes, MA was removed following detergent solubilization of the viral membrane. In contrast, in revertant preparations, the majority of MA cosedimented with the detergent-resistant complex. These results suggest that the 20LK/73EK/82AT mutations induced a significant alteration in MA-MA or MA-core interactions.

Efavirenz Is a Potent Nonnucleoside Reverse Transcriptase Inhibitor of HIV Type 1 Replication in Microgliain Vitro

2000 ◽  
Vol 16 (15) ◽  
pp. 1527-1537 ◽  
Author(s):  
Andrew V. Albright ◽  
Susan Erickson-Viitanen ◽  
Michael O'Connor ◽  
Ian Frank ◽  
Marlene M. Rayner ◽  
...  
Keyword(s):  
Reverse Transcriptase ◽  
Transcriptase Inhibitor ◽  
Nonnucleoside Reverse Transcriptase Inhibitor ◽  
Hiv Type 1 ◽  
Reverse Transcriptase Inhibitor

A Method for Combined Immunoaffinity Purification and Assay of HIV-1 Reverse Transcriptase Activity Useful for Crude Samples

1996 ◽  
Vol 235 (2) ◽  
pp. 141-152 ◽  
Author(s):  
J. Lennerstrand ◽  
A.-S. Rytting ◽  
C. Örvell ◽  
J.S. Gronowitz ◽  
C.F.R. Källander
Keyword(s):  
Reverse Transcriptase ◽  
Reverse Transcriptase Activity ◽  
Transcriptase Activity ◽  
Immunoaffinity Purification ◽  
Hiv 1
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