1174305128887S
Throughout years one to five, Dr. Elizabeth CampbellAca'a"cs group will be responsible for experiments described in Aim 2. Dr. Campbell has expertise in structural and functional studies of Mycobacterium tuberculosis transcription factors with RNA polymerase. Therefore, Dr. Campbell will lead the biochemical and structural investigations to determine the mechanisms of action of the resilience factors, ResR, WhiB2, and WhiA on transcription. The biochemical approaches to purify and assess the effect of each regulatory factor on transcriptional activity are well established in her lab. Dr. CampbellAca'a"cs group will also be using cryo-electron microscopy approaches they established to study the molecular and structural basis of how each factor coordinates with RNA polymerase to regulate resilience-associated transcription on target promoters. Her group will be using the microscopes in the Evelyn Guss Lipper Cryo-Electron Microscopy Center here at The Rockefeller University and the New York Structural Biology Center to collect data and resolve the structures to facilitate the goals of this grant. In the first year, Dr. CampbellAca'a"cs group will purify and biochemically evaluate the effects of WhiA and WhiB2 on Mtb co-regulated and independently-regulated promoters. In that period, they will also perform similar studies involving WhiB2 in the presence of reactive oxygen species, nitric oxide donors, or under anaerobic conditions to test the effects of oxidative stress and redox state on its activity. In the first year and second, they will also begin to solve the structures of these factors on their activated target promoters. In the third and fourth years, they will continue these studies with WhiA and WhiB2 on promoters independently regulated by each factor. In the fourth and fifth years, Dr. Campbell will begin biochemical studies of ResR with RNA polymerase on target sites and proceed to solve structures of ResR in complex with RNA polymerase on regulated promoters. Dr. Campbell will also perform structure-guided mutagenesis/function studies throughout all five years. Dr. Campbell will also work with Drs. Fortune and Ehrt to coordinate disseminating their findings by conference attendance and published manuscripts. The Rockefeller University
Project Grant R01AI184469 $469.9k 6/16/26 1174305128885S
The laboratory directed by Dr. Sabine EhrtAca'a"cs laboratory will identify the regulators networks of M. tuberculosis resilience variants in WhiA and WhiB2 and determine the consequences of resilience mutations on responses to antibiotics and host relevant stressors. Weill Medical College Of Cornell University
Project Grant R01AI184469 $435.5k 6/16/26 1174305128885S
The laboratory directed by Dr. Sabine EhrtAca'a"cs laboratory will identify the regulators networks of M. tuberculosis resilience variants in WhiA and WhiB2 and determine the consequences of resilience mutations on responses to antibiotics and host relevant stressors. Weill Medical College Of Cornell University
Project Grant R01AI184469 $188.1k 7/17/24 1174305128887S
Throughout years one to five, Dr. Elizabeth CampbellAca'a"cs group will be responsible for experiments described in Aim 2. Dr. Campbell has expertise in structural and functional studies of Mycobacterium tuberculosis transcription factors with RNA polymerase. Therefore, Dr. Campbell will lead the biochemical and structural investigations to determine the mechanisms of action of the resilience factors, ResR, WhiB2, and WhiA on transcription. The biochemical approaches to purify and assess the effect of each regulatory factor on transcriptional activity are well established in her lab. Dr. CampbellAca'a"cs group will also be using cryo-electron microscopy approaches they established to study the molecular and structural basis of how each factor coordinates with RNA polymerase to regulate resilience-associated transcription on target promoters. Her group will be using the microscopes in the Evelyn Guss Lipper Cryo-Electron Microscopy Center here at The Rockefeller University and the New York Structural Biology Center to collect data and resolve the structures to facilitate the goals of this grant. In the first year, Dr. CampbellAca'a"cs group will purify and biochemically evaluate the effects of WhiA and WhiB2 on Mtb co-regulated and independently-regulated promoters. In that period, they will also perform similar studies involving WhiB2 in the presence of reactive oxygen species, nitric oxide donors, or under anaerobic conditions to test the effects of oxidative stress and redox state on its activity. In the first year and second, they will also begin to solve the structures of these factors on their activated target promoters. In the third and fourth years, they will continue these studies with WhiA and WhiB2 on promoters independently regulated by each factor. In the fourth and fifth years, Dr. Campbell will begin biochemical studies of ResR with RNA polymerase on target sites and proceed to solve structures of ResR in complex with RNA polymerase on regulated promoters. Dr. Campbell will also perform structure-guided mutagenesis/function studies throughout all five years. Dr. Campbell will also work with Drs. Fortune and Ehrt to coordinate disseminating their findings by conference attendance and published manuscripts. The Rockefeller University
Project Grant R01AI184469 $190.7k 7/17/24