Search bioRxiv⌕ Search

Biology subjects

Lee, K.-i.

Publications and source records attributed to Lee, K.-i..

2 recordsLinked to original sources

Oligomeric coiled-coil adhesins that drive chain-like adhesion diversify surface colonization strategies in Shiga toxin-producing Escherichia coli

Bacteria frequently colonize host and environmental surfaces under fluid flow. Chain-like adherence pattern (CLAP) is an EibG-mediated surface colonization phenotype of certain Shiga toxin-producing Escherichia coli (STEC) that lack the locus of enterocyte effacement (LEE). EibG, an immunoglobulin-binding oligomeric coiled-coil adhesin, drives CLAP, but the temporal dynamics and genetic diversity underlying chain formation remain unclear. Here, we use live-cell time-lapse imaging to show that chains arise from single cells that elongate and divide without separation. Under flow, chains resist detachment and undergo shear-dependent fragmentation at cell-cell junctions, releasing viable clonal units that disperse downstream. Comparative genomics reveals substantial diversity among EibG-related adhesins and identifies distinct lineages, including chain-like adhesins (cla) that mediate CLAP while lacking IgG binding. Screening of 1,354 LEE-negative STEC genomes from England shows that claB is present in 95.6% of strains from major LEE-negative STEC serotypes, highlighting its epidemiological prevalence. Targeted mutagenesis demonstrates that chain formation and IgG binding are mediated by distinct structural domains, revealing the modular functional architecture of these adhesins. In a mouse infection model, deletion of eibG reduced lethality, indicating that EibG contributes to virulence. Collectively, these findings establish CLAP as a dynamic, surface-associated strategy of LEE-negative STEC and reveal previously unrecognized diversification among adhesins that drive this behavior.

microbiology↗

Genome-scale dissection of phase-variable gene function in Campylobacter jejuni using a stabilized phasotype library

Phase variation (PV) enables bacterial pathogens to rapidly alter their surface structures through reversible mutations in simple sequence repeats, promoting immune evasion and environmental adaptation. In Campylobacter jejuni, the stochastic nature of PV has hindered the systematic functional analysis of phase-variable genes (PVGs). Here, we introduce PV-GenShift, a genome-scale screening platform built on a genetically stabilized library of phase-locked C. jejuni variants. By fixing the ON/OFF states of 15 PVGs, PV-GenShift enables reproducible, high-resolution analysis of phasotypes, defined as unique ON/OFF combinations across multiple PVGs, under defined selective pressures. Using models of human serum exposure, murine colonization, and chicken gut passage, we identified distinct phasotypes associated with serum resistance and with enrichment during mouse colonization, particularly involving capsular polysaccharide modifications such as O-methyl phosphoramidation and methylation. In contrast, chicken gut passage resulted in heterogeneous ON/OFF shifts without a dominant phasotype. These findings highlight the combinatorial impact of PVG expression states on bacterial adaptation and establish PV-GenShift as a broadly applicable framework for dissecting PV-driven phenotypic diversity. This approach provides a scalable strategy for exploring genotype-phenotype relationships and offers insights relevant to vaccine design and targeted therapeutics. Significance StatementPhase variation generates phenotypic diversity that enables pathogens to evade immunity and adapt to changing environments; however, its random nature has long obscured functional analysis. This study introduces PV-GenShift, a genome-scale platform that stabilizes phase-variable gene expression in Campylobacter jejuni, allowing the systematic identification of gene combinations that influence survival under selective pressures. Using PV-GenShift, we identified phasotypes associated with serum resistance and enrichment during mouse colonization, while chicken passage produced diverse but non-specific shifts. These results demonstrate how combinatorial ON/OFF states of multiple genes shape bacterial adaptation and provide a generalizable strategy for studying phase variation across pathogens, with implications for vaccine design and targeted therapeutics.

microbiology↗