title: Selective pharmacologic inhibition of a PASTA kinase increases Listeria monocytogenes susceptibility to β-lactam antibiotics.
abstract: While β-lactam antibiotics are a critical part of the antimicrobial arsenal, they are frequently compromised by various resistance mechanisms, including changes in penicillin binding proteins of the bacterial cell wall. Genetic deletion of the penicillin binding protein and serine/threonine kinase-associated protein (PASTA) kinase in methicillin-resistant Staphylococcus aureus (MRSA) has been shown to restore β-lactam susceptibility. However, the mechanism remains unclear, and whether pharmacologic inhibition would have the same effect is unknown. In this study, we found that deletion or pharmacologic inhibition of the PASTA kinase in Listeria monocytogenes by the nonselective kinase inhibitor staurosporine results in enhanced susceptibility to both aminopenicillin and cephalosporin antibiotics. Resistance to vancomycin, another class of cell wall synthesis inhibitors, or antibiotics that inhibit protein synthesis was unaffected by staurosporine treatment. Phosphorylation assays with purified kinases revealed that staurosporine selectively inhibited the PASTA kinase of L. monocytogenes (PrkA). Importantly, staurosporine did not inhibit a L. monocytogenes kinase without a PASTA domain (Lmo0618) or the PASTA kinase from MRSA (Stk1). Finally, inhibition of PrkA with a more selective kinase inhibitor, AZD5438, similarly led to sensitization of L. monocytogenes to β-lactam antibiotics. Overall, these results suggest that pharmacologic targeting of PASTA kinases can increase the efficacy of β-lactam antibiotics.
authors.affiliation: Department of Medical Microbiology and Immunology, University of Wisconsin-Madison, Madison, Wisconsin, USA.
authors.forename: Daniel A
authors.initials: DA
authors.lastname: Pensinger
authors.affiliation: Department of Medical Microbiology and Immunology, University of Wisconsin-Madison, Madison, Wisconsin, USA Department of Medicine, University of Wisconsin-Madison, Madison, Wisconsin, USA.
authors.forename: Matthew T
authors.initials: MT
authors.lastname: Aliota
authors.affiliation: Department of Medical Microbiology and Immunology, University of Wisconsin-Madison, Madison, Wisconsin, USA Department of Medicine, University of Wisconsin-Madison, Madison, Wisconsin, USA.
authors.forename: Adam J
authors.initials: AJ
authors.lastname: Schaenzer
authors.affiliation: Department of Medical Microbiology and Immunology, University of Wisconsin-Madison, Madison, Wisconsin, USA Department of Medicine, University of Wisconsin-Madison, Madison, Wisconsin, USA.
authors.forename: Kyle M
authors.initials: KM
authors.lastname: Boldon
authors.affiliation: Department of Medicine, University of Wisconsin-Madison, Madison, Wisconsin, USA.
authors.forename: Israr-ul H
authors.initials: IU
authors.lastname: Ansari
authors.affiliation: Department of Medical Microbiology and Immunology, University of Wisconsin-Madison, Madison, Wisconsin, USA.
authors.forename: William J B
authors.initials: WJ
authors.lastname: Vincent
authors.affiliation: Department of Medical Microbiology and Immunology, University of Wisconsin-Madison, Madison, Wisconsin, USA.
authors.forename: Benjamin
authors.initials: B
authors.lastname: Knight
authors.affiliation: Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, California, USA.
authors.forename: Michelle L
authors.initials: ML
authors.lastname: Reniere
authors.affiliation: Department of Medical Microbiology and Immunology, University of Wisconsin-Madison, Madison, Wisconsin, USA Department of Medicine, University of Wisconsin-Madison, Madison, Wisconsin, USA W. S. Middleton Memorial Veterans Hospital, Madison, Wisconsin, USA.
authors.forename: Rob
authors.initials: R
authors.lastname: Striker
authors.affiliation: Department of Medical Microbiology and Immunology, University of Wisconsin-Madison, Madison, Wisconsin, USA sauer3@wisc.edu.
authors.forename: John-Demian
authors.initials: JD
authors.lastname: Sauer
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D002473:Cell Wall / Q000187:drug effects / Q000378:metabolism; D002511:Cephalosporins / Q000494:pharmacology; D024901:Drug Resistance, Multiple, Bacterial / Q000187:drug effects*; D017353:Gene Deletion; D007093:Imidazoles / Q000494:pharmacology; D008089:Listeria monocytogenes / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D046915:Penicillin-Binding Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D010406:Penicillins / Q000494:pharmacology; D010766:Phosphorylation; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D011743:Pyrimidines / Q000494:pharmacology; D019311:Staurosporine / Q000494:pharmacology; D014640:Vancomycin / Q000494:pharmacology
chemical_list: D000900:Anti-Bacterial Agents; D002511:Cephalosporins; D007093:Imidazoles; D046915:Penicillin-Binding Proteins; D010406:Penicillins; D047428:Protein Kinase Inhibitors; D011743:Pyrimidines; C521840:AZD5438; D014640:Vancomycin; D017346:Protein Serine-Threonine Kinases; D019311:Staurosporine