bioRxiv Science⌕ Search

bioRxiv · 10.1101/2025.04.19.648921

Intraspecific bacterial competition mediated by rapidly diversifying tailocin and prophage loci

Abstract

Signatures of selection in microbial genomes are often linked to biotic interactions, notably resistance to host immunity or bacteriophage attack. Here, we highlight the importance of competitive interactions, specifically between conspecific bacteria, in shaping microbial genomes, using animal-associated Xenorhabdus bacteria. An aspect of microbial genome variation is the presence of diverse mobile genetic elements that distinguish bacterial genomes from their closely related kin. We found that compared to those across domain Bacteria, Xenorhabdus genomes contain among the highest proportion of phage-related genes, and that variation among strains in their total number of protein-coding genes is largely predicted by variation in total number of non-cargo phage genes per genome. A universal yet highly variable Xenorhabdus phage-related region encodes xenorhabdicin tailocins. This region ranged in length from 12 to 41 kilobases, and its specificity-determining main tail fiber varied from 341 to 1035 amino acids. Concomitant with this variation, tailocins produced by six strains of X. nematophila differed dramatically in particle length and killing profile. Intriguingly, while X. nematophila xenorhabdicins displayed common heterospecific killing activity, they varied in conspecific killing activity. We further demonstrate the ecological importance of xenorhabdicin diversity by associating intraspecific variation in killing profiles of mitomycin-induced lysates from 42 sympatric X. bovienii strains with genes from both xenorhabdicin-encoding loci and prophages. The susceptibility profiles of strains to lysates were associated with O-antigen biosynthesis genes. Overall, our data demonstrate that through bacteriophage-mediated genome diversification, an animal-associated bacterium can tailor its weaponry and defense systems to target the closest of relatives. IMPORTANCEMicrobes exist in complex communities with each other and with animal or plant hosts. Such biotic interactions can impose strong and variable selection on microbes and are predicted to foster diversity within lineages. By examining genomes from one bacterial genus, we show that phage-associated genes are responsible for an unusually high degree of variation in protein-coding gene content. Moreover, we link this variation to functional diversity in competitive interactions among Xenorhabdus strains. Different strains of Xenorhabdus bacteria frequently co-infect an insect, which simultaneously promotes strong selection for competitive dominance within an insect host as well as opportunities for horizontal gene flow. Additionally, genomic rearrangement and homologous recombination can provide phenotypic variation for selection. The tractability of Xenorhabdus for both lab and environmental studies make them powerful for understanding microbial genome evolution and competition and this study reveals the dominant role of bacteriophage and bacteriophage-like elements in these processes.

Explore related subjects

Keep this discovery

Explore connections, maps & timelines

BibTeXRIS

Kauffman, S. J., Awori, R. M., Allwell, E. C., Taylor, A., Bashey, F., Goodrich-Blair, H.. 2025-04-19. Intraspecific bacterial competition mediated by rapidly diversifying tailocin and prophage loci. https://doi.org/10.1101/2025.04.19.648921

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related preprints

A conserved cysteine-histidine-glutamate metal site identifies DUF501 (Rv1025), an essential uncharacterised protein family of Mycobacterium tuberculosis, as a candidate metalloenzyme and drug target

A substantial fraction of the Mycobacterium tuberculosis proteome remains functionally uncharacterised. Rv1025, a 155-residue protein carrying the domain of unknown function DUF501 (Pfam PF04417), is essential by transposon mutagenesis and vulnerable by CRISPR interference, an attractive but neglected drug target, yet has never been functionally described. The family (4,370 proteins, no Gene Ontology term, no solved structure) is uncharacterised across all organisms and essential in three Actinobacterial genera. A Foldseek search of the AlphaFold model against complete structural databases finds no significant homolog, indicating a novel fold. The operon eno-divIC-Rv1025-ppx2 is conserved across the Actinobacteria phylum, yet AlphaFold-Multimer finds no direct complex between Rv1025 and its neighbour DivIC. Instead, conservation across 8,700 homologous sequences reveals a near-invariant Cys113-His115-Glu59 cluster forming a pocket. Holo AlphaFold3 predictions with Zn, Fe and Mn confidently place a divalent metal on this triad at 2.25-2.47 A; mutating the triad relocates the metal, and an independent backbone-geometry predictor recovers the same site, confirming specificity. The triad is universal across the family: present in all 1,472 near-complete bacterial sequences of the Pfam alignment, with no non-conservative substitution among the 2,228 sequences examined, a defining feature of bacterial DUF501 rather than a mycobacterial peculiarity. We propose that DUF501 is a metal-binding protein and candidate metalloenzyme, the first functional hypothesis for this family, whose conserved, essential metal pocket is a promising drug target. As the predictions build on a conservation-defined site within a fully computational study, they are supportive rather than proof of metal occupancy and warrant experimental validation.

microbiology↗

Mycoplasmal endosymbionts of Trichomonas vaginalis are associated with reduced risk for Chlamydia trachomatis endometrial infection in asymptomatic, coinfected, women.

Trichomonas vaginalis is a protozoan parasite that causes trichomoniasis, the most common curable non-viral sexually transmitted infection, and Chlamydia trachomatis is a bacterial pathogen that can ascend to the upper genital tract and cause pelvic inflammatory disease, infertility, and ectopic pregnancy. T. vaginalis harbors bacterial endosymbionts, including Candidatus Malacoplasma girerdii, an obligate symbiont, and Metamycoplasma hominis, which can live freely or symbiotically. In a 16S rRNA sequencing study of the cervicovaginal microbiome of women at high risk for chlamydial infection, Ca. M. girerdii abundance was one of 13 features predicting lack of chlamydial spread to the endometrium, despite no direct association between T. vaginalis infection and reduced chlamydial ascension. Investigating the relationship between these microorganisms further, we found that T. vaginalis vaginal abundance correlated positively with chlamydial burden in women whose infection was confined to the cervix, while a nonsignificant inverse relationship was seen in women with endometrial spread. Among participants with high chlamydial burden, Ca. M. girerdii was detected exclusively in women without endometrial infection. Both endosymbionts trended toward more frequent detection, and higher abundance, in coinfected women without endometrial spread, while M. hominis abundance correlated strongly with T. vaginalis burden in this group. These findings suggest that mycoplasmal endosymbionts of T. vaginalis, rather than T. vaginalis itself, are microbial factors limiting chlamydial ascension, and point to a three-way interaction between parasite, endosymbiont, and bacterial pathogen that shapes upper genital tract C. trachomatis infection risk.

microbiology↗

Understanding the physiological alterations of Vibrio cholerae upon exposure to L-ascorbic acid

The scourge of cholera remains a major global public health threat. It affects up to 4 million people worldwide and causes tens of thousands of deaths each year. The disease is experiencing a concerning resurgence in many parts of Africa, the Middle East, and Asia. To effectively tackle cholera and circumvent rising antimicrobial resistance, targeted biological and preventive approaches, complementing traditional rehydration, are urgently needed. In this regard, our group has demonstrated the efficacy of L-ascorbic acid in controlling the growth and pathogenesis of Vibrio cholerae in vitro. The present work further provides a mechanistic elucidation of the L-ascorbic acid-mediated physiological changes in V. cholerae and also bolsters such a non-antibiotic approach to control cholera.

microbiology↗