Research Projects:

• We are using site-directed mutagenesis of recombinant HIV-1 RT expressed in E. coli to define and study amino acid residues involved in protein stability and subunit dimerization of the HIV-1 RT.  We now have a collection of over 500 single amino acid substitutions mutants in the HIV-1 RT.  One goal of this work is to provide new insights into designing novel therapeutic agents to inhibit HIV-1 replication.

• We are studying the evolution of critical interactions involved in protein stability we identified in the HIV-1 RT.  We are using mutagenesis and molecular modeling of retroviral RTs to investigate how critical interactions for protein stability are maintained, gained, and lost during evolution.

• We are using phenotype data from our collection of HIV-1 RT mutants to evaluate amino acid substitutions in the RT of clinical HIV-1 isolates.  This analysis can serve as a method to predict if reported sequences of HIV-1 isolates are biologically functional.  The mutational data can also be used to aid in the understanding of drug resistant HIV-1 RT mutants.

• Clyde A. Hutchison III (Principal Investigator)

• John Wrobel (Research Associate)

• Michael Conrad (Research Associate)

• Ron Swanstrom (Dept of Biochemistry and Biophysics, University of North Carolina)

• Gary Pielak (Dept of Chemistry, University of North Carolina)

• Current list of our collection of HIV-1 RT mutants

• Query the Hutchison Lab mutant collection

• HIV-1 RT phenotype data [supplement to Wrobel et al. (1998), PNAS 95, 638-645]

• List of retroviruses used in our molecular modeling study of retroviral RTs

• RT sequence alignment used in our analysis of clinical HIV-1 isolates [supplement to Wrobel et al. (2000), ARHR]

• List of RT crystal structures

• HIV-1 RT amino acid sequence

• RT mutant ticker

• Reverse Transcriptase sequences in fastA format

• List of infectious HIV-1 molecular clones

Wrobel, J.A., Conrad, M.J., Bloedon, E., Swanstrom, R., and Hutchison, C.A. III  (2000).  Analysis of HIV type 1 reverse transcriptase:  Comparing sequences of viral isolates with mutational data.  AIDS Research and Human Retroviruses 16, 2049-2054.  [PubMed]

Wrobel, J.A., Chao, S.-F., Conrad, M.J., Merker, J.D., Swanstrom, R., Pielak, G.J.,  and Hutchison, C.A. III  (1998).  A genetic approach for identifying critical residues in the fingers and palm subdomains of HIV-1 reverse transcriptase.  Proc. Natl. Acad. Sci. USA 95, 638-645. [PubMed]

Chao, S.-F., Chan, V.L., Juranka, P., Kaplan, A.H., Swanstrom, R., and Hutchison, C.A. III (1995).  Mutational sensitivity patterns define critical residues in the HIV-1 reverse transcriptase palm domain.  Nucl. Acids Res. 12, 803-810.  [PubMed]

Loeb, D.D., Swanstrom, R., Everitt, L., Manchester, M., Stamper, S.E., and Hutchison, C.A. III  (1989).  Complete mutagenesis of the HIV-1 protease.  Nature 340, 397-400.  [PubMed]

Loeb, D.D., Hutchison, C.A. III, Edgell, M.H., Farmerie, W.G., and Swanstrom, R.  (1989).  Mutational analysis of human immunodeficiency virus type 1 protease suggests functional homology with aspartic proteinases.  Journal of Virology 63, 111-121.  [PubMed]

Farmerie, W.G., Loeb, D.D., Casavant, N.C., Hutchison, C.A. III, Edgell, M.H., and Swanstrom, R. (1987).  Expression and processing of the AIDS virus reverse transcriptase in E. coli.  Science 236, 305-308. [PubMed]

• HIV Sequence Database (Los Alamos)

• Stanford HIV RT and Protease Sequence Database

• Protein Data Bank

• ExPASy (Swiss Institute of Bioinformatics)

• NCBI

Click here to view more interactive Reverse Transcriptase images

Maintained by John Wrobel.