Activity-based selection of HIV-1 reverse transcriptase variants with decreased polymerization fidelity
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Sascha N. Stumpp
Abstract
HIV-1 reverse transcriptase (HIV-1 RT) copies the RNA genome of HIV-1 into DNA, thereby committing errors at an exceptionally high frequency. Viral offspring evolve rapidly and consequently are capable of evading the immune response as well as antiviral treatment. However, error-prone viral replication could drive HIV close to extinction owing to an intolerable load of deleterious mutations. We applied a genetic selection scheme to identify variants of HIV-1 RT with a further increased error rate to study the relationship between error rate and viral replication. Using this approach, we identified 16 mutator candidates, two of which were purified and further studied in vitro. One of these variant enzymes showed a generally increased mutation frequency as compared with the reference enzyme. A single amino acid residue, R448, is probably responsible for the observed effect. Mutation of this residue, which is located within the RNase H domain of HIV-1 RT, seems to perturb the interaction with template RNA and consequently affects polymerase activity and fidelity.
©2010 by Walter de Gruyter Berlin New York
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Articles in the same Issue
- Guest Editorial
- Highlight: Molecular Neurobiology
- HIGHLIGHT: MOLECULAR NEUROBIOLOGY
- Actin-mediated gene expression in neurons: the MRTF-SRF connection
- RIM proteins and their role in synapse function
- Brain tumor stem cells
- The mitochondria permeability transition pore complex in the brain with interacting proteins – promising targets for protection in neurodegenerative diseases
- Involvement of the calcium sensor GCAP1 in hereditary cone dystrophies
- Live cell imaging of cytoskeletal dynamics in neurons using fluorescence photoactivation
- REVIEWS
- Contributions of the Ah receptor to bilirubin homeostasis and its antioxidative and atheroprotective functions
- Cathepsin L in metastatic bone disease: therapeutic implications
- GENES AND NUCLEIC ACIDS
- Activity-based selection of HIV-1 reverse transcriptase variants with decreased polymerization fidelity
- CELL BIOLOGY AND SIGNALING
- Multiple protective functions of catalase against intercellular apoptosis-inducing ROS signaling of human tumor cells
- Molecular characterisation of ‘transmembrane protein 192’ (TMEM192), a novel protein of the lysosomal membrane
- PROTEOLYSIS
- Functional study of elafin cleaved by Pseudomonas aeruginosa metalloproteinases
