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Molecular mechanisms of HipA-mediated multidrug tolerance and its neutralization by HipB
Maria A Schumacher1, Kevin M Piro, Weijun Xu
1Department of Biochemistry and Molecular Biology, University of Texas, M. D. Anderson Cancer Center, Unit 1000, Houston, TX 77030, USA. maschuma@mdanderson.org
Abstract:
Bacterial multidrug tolerance is largely responsible for the inability of antibiotics to eradicate infections and is caused by a small population of dormant bacteria called persisters. HipA is a critical Escherichia coli persistence factor that is normally neutralized by HipB, a transcription repressor, which also regulates hipBA expression. Here, we report multiple structures of HipA and a HipA-HipB-DNA complex. HipA has a eukaryotic serine/threonine kinase-like fold and can phosphorylate the translation factor EF-Tu, suggesting a persistence mechanism via cell stasis. The HipA-HipB-DNA structure reveals the HipB-operator binding mechanism, approximately 70 degrees DNA bending, and unexpected HipA-DNA contacts. Dimeric HipB interacts with two HipA molecules to inhibit its kinase activity through sequestration and conformational inactivation. Combined, these studies suggest mechanisms for HipA-mediated persistence and its neutralization by HipB.
Insights
Bacterial multidrug tolerance, driven by persister cells, is a major antibiotic challenge. Researchers elucidated the structure and function of HipA, a key persistence factor, and its inhibitor HipB, revealing mechanisms of antibiotic resistance.
Area of Science:
- Microbiology
- Molecular Biology
- Structural Biology
Background:
- Bacterial multidrug tolerance hinders antibiotic efficacy.
- Persister cells, a dormant subpopulation, are responsible for this tolerance.
- The HipA protein in Escherichia coli is a critical factor in bacterial persistence.
Purpose of the Study:
- To elucidate the structural basis of HipA's kinase activity.
- To understand the mechanism by which HipB neutralizes HipA.
- To reveal the molecular interactions governing HipA-mediated persistence and HipB regulation.
Main Methods:
- X-ray crystallography was used to determine the structures of HipA and a HipA-HipB-DNA complex.
- Biochemical assays were performed to assess HipA's kinase activity and HipB's inhibitory function.
Main Results:
- HipA exhibits a eukaryotic serine/threonine kinase-like fold and phosphorylates the translation factor EF-Tu, suggesting a role in cell stasis.
- The HipA-HipB-DNA complex structure revealed HipB's DNA-binding mechanism, significant DNA bending, and novel HipA-DNA interactions.
- Dimeric HipB inhibits HipA's kinase activity through sequestration and conformational inactivation.
Conclusions:
- These findings provide insights into the molecular mechanisms of HipA-mediated bacterial persistence.
- The study reveals how HipB counteracts HipA activity, offering potential targets for antibiotic development.
- Understanding these interactions is crucial for overcoming antibiotic resistance.
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