Nicolas Sluis-Cremer, PhD, has been awarded funding of $3,708,180 for a five-year R01 grant by the National Institutes of Health/National Institute of Allergy and Infectious Diseases (NIH/NIAID) entitled “‘On the Fly’ Time Resolved Cryo-EM Studies of Intermediate HIV-1 RT Transition States.” This proposal was submitted in response to the competitive funding announcement entitled “NIH Research Progress Grant (Parent R01 Clinical Trial Not Allowed)” under funding opportunity number PA-20-185.
Historically, interactions between substrates and/or inhibitors and enzymes have been viewed in terms of binding of small molecules to relatively rigid protein targets. However, computational and experimental studies have revealed that many proteins, in particular DNA polymerases, undergo molecular motions over a wide range of timescales. Such conformational flexibility is critical for enzymatic activity, drug action, and drug resistance. Moreover, contemporary structural biology approaches, such as X-ray crystallography, only have the ability to resolve the structures of thermodynamically stable species, and cannot inform on kinetic intermediates.
Nic, along with Dr. Guillermo Calero (Department of Structural Biology) and Dr. Alexander Deiters (Chemistry) will serve as MPIs in this research. The team will further develop and use cutting-edge technology – “on-the-fly” time-resolved cryo-electron microscopy – that, for the first time, facilitates visualization of novel protein conformations, including those transiently occurring during catalysis. They will apply this technology to address biologically and clinically relevant knowledge gaps in HIV-1 reverse transcriptase biology, with specific focus on the incorporation of nucleosides and nucleoside inhibitors, nucleoside inhibitor resistance, and the relationship between DNA synthesis and ribonuclease H activity.