Bacteria use energy dependent proteases to respond to stressful conditions. These proteases serve a dual role: destroying aberrant, potentially toxic, damaged proteins and generating stress responsive signals through degradation of regulatory factors. Cells often arrest replication in response to stress, but how regulated proteolysis contributes to cell cycle arrest in bacteria is currently poorly understood. This proposal addresses how stress related proteases target replication factors using a combination of biochemical, genetic and proteomic approaches, specifically focusing on proteases and replication factors from the model bacteria Caulobacter crescentus.
Aims 1 and 2 determine how misfolded proteins generated during proteotoxic stress directly stimulate the Lon protease to destroy the replication initiator DnaA and cause growth arrest during stress.
Aims 3 and 4 focus on how partial processing of the clamp loader subunit DnaX by the ClpXP protease is critical for replication stress tolerance during DNA damage. Because these proteases and replication factors are highly conserved throughout all bacteria, these results will impact our general understanding of replication, proteolysis, and stress tolerance. The critical role of these proteases in bacterial virulence and pathogenicity, together with the universal requirement for these proteases in bacterial stress responses, suggests that they are excellent targets for development of new antibiotic strategies that are of immediate human health need.

Public Health Relevance

Energy dependent proteases are found in all bacteria and necessary for virulence in many human pathogens. When bacteria are stressed, such as when they invade a host or encounter antibiotics, a crucial response is to stop growing in order to repair damages before they invest limited resources in growth. By determining how these changes rely on proteolytic degradation of essential replication factors, this work will reveal new path- ways that could be targeted to block bacterial virulence or to prevent bacteria from resisting the stresses produced by currently used antibiotics.

Agency
National Institute of Health (NIH)
Type
Research Project (R01)
Project #
1R01GM111706-01
Application #
8760229
Study Section
Prokaryotic Cell and Molecular Biology Study Section (PCMB)
Program Officer
Reddy, Michael K
Project Start
Project End
Budget Start
Budget End
Support Year
1
Fiscal Year
2014
Total Cost
Indirect Cost
Name
University of Massachusetts Amherst
Department
Biochemistry
Type
Schools of Arts and Sciences
DUNS #
City
Amherst
State
MA
Country
United States
Zip Code
01003
Liu, Jing; Francis, Laura I; Jonas, Kristina et al. (2016) ClpAP is an auxiliary protease for DnaA degradation in Caulobacter crescentus. Mol Microbiol 102:1075-1085
Vass, Robert H; Zeinert, Rilee D; Chien, Peter (2016) Protease regulation and capacity during Caulobacter growth. Curr Opin Microbiol 34:75-81
Vass, Robert H; Chien, Peter (2016) Two ways to skin a cat: acyldepsipeptides antibiotics can kill bacteria through activation or inhibition of ClpP activity. Mol Microbiol 101:183-5
Joshi, Kamal Kishore; Bergé, Matthieu; Radhakrishnan, Sunish Kumar et al. (2015) An Adaptor Hierarchy Regulates Proteolysis during a Bacterial Cell Cycle. Cell 163:419-31
Lau, Joanne; Hernandez-Alicea, Lisa; Vass, Robert H et al. (2015) A Phosphosignaling Adaptor Primes the AAA+ Protease ClpXP to Drive Cell Cycle-Regulated Proteolysis. Mol Cell 59:104-16
Mukherjee, Sampriti; Bree, Anna C; Liu, Jing et al. (2015) Adaptor-mediated Lon proteolysis restricts Bacillus subtilis hyperflagellation. Proc Natl Acad Sci U S A 112:250-5
Schallies, Karla B; Sadowski, Craig; Meng, Julia et al. (2015) Sinorhizobium meliloti CtrA Stability Is Regulated in a CbrA-Dependent Manner That Is Influenced by CpdR1. J Bacteriol 197:2139-49
Chien, Peter (2015) Not Throwing Baby Out with the Bathwater. Plant Cell 27:2669-70
Liu, Jing; Chien, Peter (2014) Structure and activation of a heteromeric protease complex. Proc Natl Acad Sci U S A 111:15289-90
Smith, Stephen C; Joshi, Kamal K; Zik, Justin J et al. (2014) Cell cycle-dependent adaptor complex for ClpXP-mediated proteolysis directly integrates phosphorylation and second messenger signals. Proc Natl Acad Sci U S A 111:14229-34

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