Regulation of mycobacterial adaptive mistranslation
Ended Project Grant Awarded Last action
- Award ID
- R56AI185094
- Recipient
-
The Regents Of The University Of California. San Francisco UEI KMH5K9V7S518
- Awarding agency
-
National Institute of Allergy and Infectious Diseases
National Institutes of Health, Department of Health and Human Services
- Obligated
- $1,359,316
Award facts
- Assistance listing
- 93.855 Allergy and Infectious Diseases Research
- Completion date
- Originally
- Place of performance
- San Francisco, CA 94110, USA
Award details come from USAspending.gov.
Description
- Abstract
- Mycobacterium tuberculosis (MTB), the causative agent of tuberculosis (TB) uses regulation of specific translation error rates - mistranslation - to survive a variety of stressors including antibiotics and the host environment. Both too high and too low mistranslation rates can be detrimental to the pathogen. However, IT is not known how MTB regulates translational error. Understanding the regulation of mistranslation by MTB will allow for the development of targeted therapies that interfere with MTB's tolerance to antibiotics and survival within the host. The source of adaptive mistranslation in MTB is the indirect trna aminoacylation pathway: used for the cognate synthesis of aminoacylated glutamine and asparagine trnas, and which results in high levels of mischarged glutamine and asparagine trnas. This pathway is present in most pathogens with the exception of escherichia coli. Using forward genetics and small molecule screens, we have identified drugs and genetic mutations that are implicated in MTB's regulation of high mistranslation rates. The aminoglycoside kasugamycin (KSG) can decrease specific mistranslation rates in MTB but has very limited anti-microbial activity and has a very high minimum inhibitory concentration (MIC) against MTB growth in vitro. When given at submic concentrations to MTB-infected mice, KSG can, as a single agent, attenuate MTB survival; and when given in combination, can substantially potentiate anti-TB drugs. Similarly, deletion of the 16S rrna methyltransferase gidb allows low/moderate levels of mistranslation but prevents runaway catastrophic mistranslation. Both of these perturbations affect the ribosome. This is intriguing because prior studies on ribosomes had suggested that ribosomes do not discriminate against mischarged trnas. A potential explanation may be that those prior studies were performed exclusively with ribosomes from e. Coli - which, unlike MTB, doesn't routinely encounter mischarged trnas. To understand the mechanisms by which the MTB translational apparatus can discriminate against mischarged trnas we will use biochemical, structural and kinetic approaches. We will solve the structure of MTB ribosomes +/- KSG or +/- gidb-mediated methylation to identify how these ribosomal perturbations affect ribosomal recognition of, and discrimination of mischarged trnas. Using single molecule fluorescence resonance energy transfer (smfret), we will identify the steps in the translation cycle in which ribosomes can proofread against mischarged trna-mediated mistranslation. Together, these studies will a) allow mechanistic insight into an important non-genetic and non-transcriptional mechanism by which MTB survives diverse stressors, b) provide greater understanding of MTB ribosome function and the translation cycle, which is likely to be substantially different from model organisms, and c) permit the future development of structure- and biochemical-informed therapies that target MTB adaptive mistranslation.
Transactions
| Action | Obligated | Date |
|---|---|---|
| Amendment | $0 | |
| Amendment | $679,658 | |
| Original award | $679,658 |
Subawards
| Subrecipient | Amount | Awarded |
|---|---|---|
| The Trustees Of Columbia University In The City Of New York | $65,000 |
Quick answers
Who received award R56AI185094?
R56AI185094 was awarded to The Regents Of The University Of California. San Francisco by National Institute of Allergy and Infectious Diseases (National Institutes of Health) on August 16, 2024.
How much is award R56AI185094 worth?
$1,359,316 has been obligated on R56AI185094.
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