The focus of our research is to determine how signal transduction regulates directed motility and behavior in the bacterium Myxococcus xanthus using an integrated approach that combines biochemistry, genetics, and cell biology. M. xanthus is an excellent model system to address fundamental questions concerning cell-cell signaling and directed movement as cells form multicellular biofilms and fruiting bodies as part of a complex life cycle. M. xanthus fruiting bodies are similar in many ways to biofilms formed by some pathogenic bacteria and are of public health interest since biofilms render bacteria resistant to antibiotics and are very difficult to treat in patients. Biofilm and fruiting body formation require the activity of chemosensory systems to direct cell movements. Previously, we have shown that the Frz chemosensory pathway regulates both vegetative swarming and developmental aggregation by controlling the reversal frequency of cells. Cell reversals in Myxococcus, like tumbling in flagellated bacteria, allows cells to reorient themselves and to bias directional motility based on the temporal sensing of stimuli. In the last grant period, our studie of the Frz system have allowed us to identify important proteins responsible for the functioning of the gliding motility engines. Our first specific aim is to continue our characterization of the dynamics of the motor protein AglR, a MotA homolog, at single molecule resolution. For these experiments, we are tracking the AglR motility complexes with photoactivatable localization microscopy (PALM) at millisecond intervals. These experiments should provide important information on this novel motility system. The techniques being utilized are innovative because they permit analysis of the dynamics of single molecule motors at nanometer resolution. Our second specific aim is to finalize our study of the Frz pathway by identifying the output of the pathway: the proteins that interact directly with the phosphorylated receiver domain, FrzZ. The output of the pathway is responsible for transmitting information from the Frz chemosensory pathway to the motors and the controllers of cell polarity, reversing the gliding motility and Type IV pili engines of M. xanthus. These experiments are significant because the coupling between the chemosensory system and the gliding motility engines is very different from the E. coli paradigm and likely to be utilized by other bacterial systems.

Public Health Relevance

The focus of our research is to understand how signal transduction regulates motility and multicellular interactions in the bacterium Myxococcus xanthus. Myxococcus forms fruiting bodies that are similar in many ways to biofilms formed by Pseudomonas aeruginosa and other pathogens;these are of public health interest since biofilms render bacteria resistant to antibiotics and are very difficult to treat in patients. Bioflm formation usually requires motility, chemotaxis, type IV pili and extracellular polysaccharide matrix materials;these are more easily studied in Myxococcus, a non-pathogenic bacterium, but the results are directly relevant to the understanding and control of pathogenic bacteria with similar properties.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
2R01GM020509-40
Application #
8577971
Study Section
Prokaryotic Cell and Molecular Biology Study Section (PCMB)
Program Officer
Reddy, Michael K
Project Start
1976-06-01
Project End
2015-08-31
Budget Start
2013-09-01
Budget End
2014-08-31
Support Year
40
Fiscal Year
2013
Total Cost
$352,125
Indirect Cost
$127,125
Name
University of California Berkeley
Department
Biochemistry
Type
Schools of Arts and Sciences
DUNS #
124726725
City
Berkeley
State
CA
Country
United States
Zip Code
94704
Moine, Audrey; Agrebi, Rym; Espinosa, Leon et al. (2014) Functional organization of a multimodular bacterial chemosensory apparatus. PLoS Genet 10:e1004164
Nan, Beiyan; McBride, Mark J; Chen, Jing et al. (2014) Bacteria that glide with helical tracks. Curr Biol 24:R169-73
Kaimer, Christine; Zusman, David R (2013) Phosphorylation-dependent localization of the response regulator FrzZ signals cell reversals in Myxococcus xanthus. Mol Microbiol 88:740-53
He, Xuesong; Tian, Yan; Guo, Lihong et al. (2010) Oral-derived bacterial flora defends its domain by recognizing and killing intruders--a molecular analysis using Escherichia coli as a model intestinal bacterium. Microb Ecol 60:655-64
Nan, Beiyan; Mauriello, Emilia M F; Sun, Im-Hong et al. (2010) A multi-protein complex from Myxococcus xanthus required for bacterial gliding motility. Mol Microbiol 76:1539-54
Scott, Ansley E; Simon, Eric; Park, Samuel K et al. (2008) Site-specific receptor methylation of FrzCD in Myxococcus xanthus is controlled by a tetra-trico peptide repeat (TPR) containing regulatory domain of the FrzF methyltransferase. Mol Microbiol 69:724-35
Inclan, Yuki F; Laurent, Sophie; Zusman, David R (2008) The receiver domain of FrzE, a CheA-CheY fusion protein, regulates the CheA histidine kinase activity and downstream signalling to the A- and S-motility systems of Myxococcus xanthus. Mol Microbiol 68:1328-39
Mauriello, Emilia Mf; Zusman, David R (2007) Polarity of motility systems in Myxococcus xanthus. Curr Opin Microbiol 10:624-9