In vivo protein interaction network analysis reveals porin-localized antibiotic inactivation in Acinetobacter baumannii strain AB5075.
The nosocomial pathogen Acinetobacter baumannii is a frequent cause of hospital-acquired infections worldwide and is a challenge for treatment due to its evolved resistance to antibiotics, including carbapenems. Here, to gain insight on A. baumannii antibiotic resistance mechanisms, we analyse the protein interaction network of a multidrug-resistant A. baumannii clinical strain (AB5075). Using in vivo chemical cross-linking and mass spectrometry, we identify 2,068 non-redundant cross-linked peptide pairs containing 245 intra- and 398 inter-molecular interactions. Outer membrane proteins OmpA and YiaD, and carbapenemase Oxa-23 are hubs of the identified interaction network. Eighteen novel interactors of Oxa-23 are identified. Interactions of Oxa-23 with outer membrane porins OmpA and CarO are verified with co-immunoprecipitation analysis. Furthermore, transposon mutagenesis of oxa-23 or interactors of Oxa-23 demonstrates changes in meropenem or imipenem sensitivity in strain AB5075. These results provide a view of porin-localized antibiotic inactivation and increase understanding of bacterial antibiotic resistance mechanisms.
Department of Genome Sciences, University of Washington, 850 Republican Street, Brotman Building Room 154, Seattle, Washington 98109, USA.
Xia
http://orcid.org/0000-0002-0024-4481
X
Wu
Department of Genome Sciences, University of Washington, 850 Republican Street, Brotman Building Room 154, Seattle, Washington 98109, USA.
Juan D
JD
Chavez
Department of Genome Sciences, University of Washington, 850 Republican Street, Brotman Building Room 154, Seattle, Washington 98109, USA.
Devin K
http://orcid.org/0000-0002-3241-6276
DK
Schweppe
Department of Chemistry, University of Washington, Seattle, Washington 98109, USA.
Chunxiang
C
Zheng
Department of Genome Sciences, University of Washington, 850 Republican Street, Brotman Building Room 154, Seattle, Washington 98109, USA.
Chad R
CR
Weisbrod
Department of Genome Sciences, University of Washington, 850 Republican Street, Brotman Building Room 154, Seattle, Washington 98109, USA.
Jimmy K
JK
Eng
Department of Genome Sciences, University of Washington, 850 Republican Street, Brotman Building Room 154, Seattle, Washington 98109, USA.
Ananya
A
Murali
Department of Genome Sciences, University of Washington, 850 Republican Street, Brotman Building Room 154, Seattle, Washington 98109, USA.
Samuel A
SA
Lee
Department of Genome Sciences, University of Washington, 850 Republican Street, Brotman Building Room 154, Seattle, Washington 98109, USA.
Elizabeth
E
Ramage
Department of Genome Sciences, University of Washington, 850 Republican Street, Brotman Building Room 154, Seattle, Washington 98109, USA.
Larry A
LA
Gallagher
Department of Microbiology, University of Washington, Seattle, Washington 98195, USA.
Hemantha D
HD
Kulasekara
Department of Microbiology, University of Washington, Seattle, Washington 98195, USA.
Mauna E
ME
Edrozo
Department of Microbiology, University of Washington, Seattle, Washington 98195, USA.
Cassandra N
CN
Kamischke
Department of Microbiology, University of Washington, Seattle, Washington 98195, USA.
Mitchell J
MJ
Brittnacher
Department of Genome Sciences, University of Washington, 850 Republican Street, Brotman Building Room 154, Seattle, Washington 98109, USA.|Department of Microbiology, University of Washington, Seattle, Washington 98195, USA.|Department of Medicine, University of Washington, Seattle, Washington 98195, USA.
Samuel I
SI
Miller
Department of Microbiology, University of Washington, Seattle, Washington 98195, USA.|Department of Medicine, University of Washington, Seattle, Washington 98195, USA.
Pradeep K
PK
Singh
Department of Genome Sciences, University of Washington, 850 Republican Street, Brotman Building Room 154, Seattle, Washington 98109, USA.
Colin
C
Manoil
Department of Genome Sciences, University of Washington, 850 Republican Street, Brotman Building Room 154, Seattle, Washington 98109, USA.|Department of Chemistry, University of Washington, Seattle, Washington 98109, USA.
James E
JE
Bruce
D040981:Acinetobacter baumannii / Q000145:classification / Q000187:drug effects* / Q000378:metabolism; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D024881:Drug Resistance, Bacterial / Q000235:genetics / Q000502:physiology*; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects / Q000502:physiology; D053263:Gene Regulatory Networks; D013058:Mass Spectrometry; D008958:Models, Molecular; D018272:Porins / Q000378:metabolism*; D011487:Protein Conformation; D000066608:Underage Drinking
D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D018272:Porins