Klebsiella pneumoniae DedA family proteins have redundant roles in divalent cation homeostasis and resistance to phagocytosis

• Published in: Microbiology spectrum Vol. 12; no. 2; p. e0380723
• Main Authors: Tiwari, Vijay, Sharma, Amit, Braga, Reygan, Garcia, Emily, Appiah, Ridhwana, Fleeman, Renee, Abuaita, Basel H, Patrauchan, Marianna, Doerrler, William T
• Format: Journal Article
• Language: English
• Published: United States American Society for Microbiology 06-02-2024
• Subjects: capsular polysaccharide; divalent cations; membrane transport; Molecular and Cellular Biology; phagocytosis; Research Article; phagocytosis; membrane transport; capsular polysaccharide; divalent cations
• ISSN: 2165-0497; 2165-0497
• DOI: 10.1128/spectrum.03807-23

The DedA superfamily is a highly conserved family of membrane proteins. Deletion of
and
, encoding related DedA family proteins, results in sensitivity to elevated temperature, antibiotics, and alkaline pH. The human pathogen
possesses genes encoding DedA family proteins with >90% amino acid identity to
YqjA and YghB. We hypothesized that the deletion of
and
will impact its physiology and may reduce its virulence. The
Δ
Δ
mutant (strain VT101) displayed a growth defect at 42°C and alkaline pH sensitivity, not unlike its
counterpart. However, VT101 retained mostly wild-type resistance to antibiotics. We found VT101 was sensitive to the chelating agent EDTA, the anionic detergent SDS, and agents capable of alkalizing the bacterial cytoplasm such as bicarbonate or chloroquine. We could restore growth at alkaline pH and at elevated temperature by addition of 0.5-2 mM Ca
or Mg
to the culture media. VT101 displayed a slower uptake of calcium, which was dependent upon calcium channel activity. VT201, with similar deletions as VT101 but derived from a virulent
strain, was highly susceptible to phagocytosis by alveolar macrophages and displayed a defect in the production of capsule. These findings suggest divalent cation homeostasis and virulence are interlinked by common functions of the DedA family.IMPORTANCE
is a dangerous human pathogen. The DedA protein family is found in all bacteria and is a membrane transporter often required for virulence and antibiotic resistance.
possesses homologs of
YqjA and YghB, with 60% amino acid identity and redundant functions, which we have previously shown to be required for tolerance to biocides and alkaline pH. A
strain lacking
and
was found to be sensitive to alkaline pH, elevated temperature, and EDTA/SDS and displayed a defect in calcium uptake. Sensitivity to these conditions was reversed by addition of calcium or magnesium to the growth medium. Introduction of Δ
and Δ
mutations into virulent
resulted in the loss of capsule, increased phagocytosis by macrophages, and a partial loss of virulence. These results show that targeting the
DedA family results in impaired divalent cation transport and, in turn, loss of virulence.