The Y271 and I274 Amino Acids in Reverse Transcriptase of Human Immunodeficiency Virus-1 Are Critical to Protein Stability
Open Access
- 3 July 2009
- journal article
- research article
- Published by Public Library of Science (PLoS) in PLOS ONE
- Vol. 4 (7), e6108
- https://doi.org/10.1371/journal.pone.0006108
Abstract
Reverse transcriptase (RT) of human immunodeficiency virus (HIV)-1 plays a key role in initiating viral replication and is an important target for developing anti-HIV drugs. Our previous study showed that two mutations (Y271A and I274A) in the turn RT (Gln269-Arg277) abrogated viral replication, but the replication capacity and RT activity was discordant. In this study, we further investigated why alanine substitutions at these two sites would affect viral replication. We found that both RT activity and RT protein were almost undetectable in viral particles of these two mutants, although the Pr160gag-pol mutants were properly expressed, transported and incorporated. Using protease inhibition assay, we demonstrated a correlation between the degradation of the RT mutants and the activity of viral protease. Our native gel analysis indicated that the mutations at 271 and 274 amino acids might cause conformational changes, leading to the formation of higher order oligomers instead of dimers, resulting in increased protein instability and susceptibility to viral protease. Thus, residues 271 and 274 are critical to RT stability and resistance to viral protease. The conservation of the two amino acid residues among different strains of HIV-1 lent further support to this conclusion. The knowledge gained here may prove useful in drug design.Keywords
This publication has 42 references indexed in Scilit:
- Mutational analysis of the “turn” of helix clamp motif of HIV-1 reverse transcriptaseBiochemical and Biophysical Research Communications, 2008
- Generation of Infectious Molecular Clones of Simian Immunodeficiency Virus from Fecal Consensus Sequences of Wild ChimpanzeesJournal of Virology, 2007
- Incorporation of Human Immunodeficiency Virus Type 1 Reverse Transcriptase into Virus-Like ParticlesJournal of Virology, 2007
- Mutational analysis revealed that conservation of hepatitis B virus reverse transcriptase residue 306 (rtP306) is crucial for encapsidation of pregenomic RNAFEBS Letters, 2007
- Severe acute respiratory syndrome-associated coronavirus 3a protein forms an ion channel and modulates virus releaseProceedings of the National Academy of Sciences of the United States of America, 2006
- Virion Instability of Human Immunodeficiency Virus Type 1 Reverse Transcriptase (RT) Mutated in the Protease Cleavage Site between RT p51 and the RT RNase H DomainJournal of Virology, 2005
- Mutations That Abrogate Human Immunodeficiency Virus Type 1 Reverse Transcriptase Dimerization Affect Maturation of the Reverse Transcriptase HeterodimerJournal of Virology, 2005
- Effects of mutations in the polymerase domain on the polymerase, RNase H and strand transfer activities of human immunodeficiency virus type 1 reverse transcriptaseJournal of Molecular Biology, 1998
- Conformational stability of dimeric HIV-1 and HIV-2 reverse transcriptasesBiochemistry, 1995
- Reduced Frameshift Fidelity and Processivity of HIV-1 Reverse Transcriptase Mutants Containing Alanine Substitutions in Helix H of the Thumb SubdomainPublished by Elsevier BV ,1995