Urine-mediated suppression of
Klebsiella pneumoniae
mucoidy is counteracted by spontaneous Wzc variants altering capsule chain length
Authors: Khadka Saroj , Ring Brooke E. , Walker Ryan S. , Krzeminski Lindsey R. , Pariseau Drew A. , Hathaway Matthew , Mobley Harry L. T. , Mike Laura A. 
Published: August 2023

Klebsiella pneumoniae
is a hospital-associated pathogen primarily causing urinary tract infections (UTIs), pneumonia, and septicemia. Two challenging lineages include the hypervirulent strains, causing invasive community-acquired infections, and the carbapenem-resistant classical strains, most frequently isolated from UTIs. While hypervirulent strains are often characterized by a hypermucoid phenotype, classical strains usually present with low mucoidy. Since clinical UTI isolates tend to exhibit limited mucoidy, we hypothesized that environmental conditions may drive
K. pneumoniae
adaptation to the urinary tract and select against mucoid isolates. We found that both hypervirulent
K. pneumoniae
and classical
Klebsiella
UTI isolates significantly suppressed mucoidy when cultured in urine without reducing capsule abundance. A genetic screen identified secondary mutations in the
wzc
tyrosine kinase that overcome urine-suppressed mucoidy. Over-expressing Wzc variants
in trans
was sufficient to boost mucoidy in both hypervirulent and classical
Klebsiella
UTI isolates. Wzc is a bacterial tyrosine kinase that regulates capsule polymerization and extrusion. Although some Wzc variants reduced Wzc phospho-status, urine did not alter Wzc phospho-status. Urine does, however, increase
K. pneumoniae
capsule chain length diversity and enhance cell-surface attachment. The identified Wzc variants counteract urine-mediated effects on capsule chain length and cell attachment. Combined, these data indicate that capsule chain length correlates with
K. pneumoniae
mucoidy and that this extracellular feature can be fine-tuned by spontaneous Wzc mutations, which alter host interactions. Spontaneous Wzc mutation represents a global mechanism that could fine-tune
K. pneumoniae
niche-specific fitness in both classical and hypervirulent isolates.

IMPORTANCE

Klebsiella pneumoniae
is high-priority pathogen causing both hospital-associated infections, such as urinary tract infections, and community-acquired infections. Clinical isolates from community-acquired infection are often characterized by a tacky, hypermucoid phenotype, while urinary tract isolates are usually not mucoid. Historically, mucoidy was attributed to capsule overproduction; however, recent reports have demonstrated that
K. pneumoniae
capsule abundance and mucoidy are not always correlated. Here, we report that human urine suppresses
K. pneumoniae
mucoidy, diversifies capsule polysaccharide chain length, and increases cell surface association. Moreover, specific mutations in the capsule biosynthesis gene,
wzc
, are sufficient to overcome urine-mediated suppression of mucoidy. These Wzc variants cause constitutive production of more uniform capsular polysaccharide chains and increased release of capsule from the cell surface, even in urine. These data demonstrate that
K. pneumoniae
regulates capsule chain length and cell surface attachment in response host cues, which can alter bacteria-host interactions.