Robert Craigie, Ph.D.

Senior Investigator

Molecular Virology Section, Laboratory of Molecular Biology

NIDDK

Building 5, Room 301
5 Memorial Drive
Bethesda, MD 20814

301-496-4081

bobc@helix.nih.gov

Research Topics

Current Research

Our research focuses on mechanistic aspects of retroviral DNA integration.  After entering the host cell, a DNA copy of the viral genome is made by reverse transcription.  Integration of this viral DNA into a chromosome of the host cell is an essential step in the retroviral replication cycle. 

The key player in the retroviral DNA (deoxyribonucleic acid) integration process is the virally encoded integrase protein.  Integrase processes the ends of the viral DNA and covalently inserts these processed ends into host DNA.  We study the molecular mechanism of these reactions using biochemical, biophysical, and structural techniques.  Close collaborations with X-ray crystallography and NMR (Nuclear Magnetic Resonance) groups in the NIDDK form an integral part of our research.

Cellular proteins play important accessory roles in the integration process.  A focus of our research on cellular factors has been the mechanism that prevents integrase using the viral DNA as a target for integration.  Such autointegration would result in destruction of the viral DNA.  We have identified a cellular protein, which we called barrier-to-autointegration factor (BAF), that prevents integration of the viral DNA into itself. BAF is a DNA bridging protein that bridges together segments of double-stranded DNA.  At high DNA concentration, this would result in aggregation.  However, at low DNA concentration, such as the few copies of viral DNA in the cytoplasm of an infected cell, the DNA bridging property of BAF results in intracellular compaction.  Our model is that compaction of the viral DNA by BAF makes it inaccessible as a target for integration. 

Applying our Research

This research will help the public with understanding the mechanism of action of HIV-1 integrase inhibitors and how the virus is able to develop resistance.

Biography

  • Ph.D., London, 1982
  • B.S., London, 1978

Selected Publications

  1. Passos DO, Li M, Yang R, Rebensburg SV, Ghirlando R, Jeon Y, Shkriabai N, Kvaratskhelia M, Craigie R, Lyumkis D. Cryo-EM structures and atomic model of the HIV-1 strand transfer complex intasome. Science. 2017;355(6320):89-92.

  2. Li M, Jurado KA, Lin S, Engelman A, Craigie R. Engineered hyperactive integrase for concerted HIV-1 DNA integration. PLoS One. 2014;9(8):e105078.

  3. Li M, Ivanov V, Mizuuchi M, Mizuuchi K, Craigie R. DNA requirements for assembly and stability of HIV-1 intasomes. Protein Sci. 2012;21(2):249-57.

  4. Li M, Lin S, Craigie R. Outer domains of integrase within retroviral intasomes are dispensible for catalysis of DNA integration. Protein Sci. 2016;25(2):472-8.

  5. Craigie R. The road to HIV-1 integrase inhibitors: the case for supporting basic research. Future Virol. 2014;9(10):899-903.


This page was last updated on February 18th, 2016