Riboswitches are structured RNAs that undergo significant conformational changes in response to specific ligands as a mechanism to regulate gene expression. In addition to several proteins, two different classes of c-di-GMP riboswitches have been implicated in the cyclic diguanosine monophosphate (c-di-GMP) second-messenger pathway, a ubiquitous signaling method in bacteria. The crystal structures for both classes of c-di-GMP riboswitches have been solved showing two different mechanisms for c-di-GMP recognition. These riboswitches are located in the 5'-untranslated regions of genes involved in biofilm formation, cell motility, and virulence factors. Since c-di-GMP riboswitches are prevalent in bacterial species responsible for tetanus, anthrax, botulism, and cholera among others, their characterization has direct therapeutic applications. Although the structure of both classes of riboswitches has been extensively characterized, very few experiments have been performed in vivo. The Strobel lab has synthesized a series of c-di-GMP analogues to differentially study the ligand binding characteristics in both c-di-GMP riboswitches. Several of these ligands also show resistance to intracellular cleavage and are ideal for studying the behavior of c-di-GMP riboswitches in the cell. This proposal seeks to determine the functions of c-di-GMP in the cell and define the role of the c-di-GMP riboswitch on bacterial gene expression using a multidisciplinary approach spanning biochemistry, structural biology, microbiology, and genetics.

Public Health Relevance

Riboswitches are structured RNAs that change their conformation in response to specific metabolites and thus alter gene expression. Two such RNA receptors that recognize cyclic diguanosine monophosphate (c- di-GMP) have been implicated in the ubiquitous c-di-GMP second-messenger pathway. These are found in the disease causative agents for anthrax, botulism, tetanus, and cholera among others. This proposal aims to study the cyclic-di-GMP riboswitches under native conditions in the cell.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Postdoctoral Individual National Research Service Award (F32)
Project #
1F32GM115097-01A1
Application #
9048361
Study Section
Special Emphasis Panel (ZRG1)
Program Officer
Lees, Robert G
Project Start
2016-04-01
Project End
2018-03-31
Budget Start
2016-04-01
Budget End
2017-03-31
Support Year
1
Fiscal Year
2016
Total Cost
Indirect Cost
Name
Yale University
Department
Biochemistry
Type
Schools of Medicine
DUNS #
043207562
City
New Haven
State
CT
Country
United States
Zip Code