Caenorhabditis elegans mutants resistant to attachment of Yersinia biofilms

Abstract

The detailed composition and structure of the Caenorhabditis elegans surface are unknown. Previous genetic studies used antibody or lectin binding to identify srf genes that play roles in surface determination. Infection by Microbacterium nematophilum identified bus (bacterially unswollen) genes that also affect surface characteristics. We report that biofilms produced by Yersinia pestis and Y. pseudotuberculosis, which bind the C. elegans surface predominantly on the head, can be used to identify additional surface-determining genes. A screen for C. elegans mutants with a biofilm absent on the head (Bah) phenotype identified three novel genes: bah-1, bah-2, and bah-3. The bah-1 and bah-2 mutants have slightly fragile cuticles but are neither Srf nor Bus, suggesting that they are specific for surface components involved in biofilm attachment. A bah-3 mutant has normal cuticle integrity, but shows a stage-specific Srf phenotype. The screen produced alleles of five known surface genes: srf-2, srf-3, bus-4, bus-12, and bus-17. For the X-linked bus-17, a paternal effect was observed in biofilm assays.

Figure 1.—

Aberrant locomotion on bacterial lawns. (A) Normal locomotion by wild-type strain N2 on E. coli OP50, the standard laboratory food. (B) Skd phenotype of N2 on Y. pseudotuberculosis. (C) Normal locomotion by bah-3(br9) on Y. pseudotuberculosis. (D) Skd phenotype of bus-17(br11) on E. coli.

Figure 2.—

Biofilm phenotype of srf-6 mutants. Biofilm presence was scored 4 hr after transfer of animals to Y. pseudotuberculosis lawns. Solid bars, L2; open bars, L3; horizontal hatching, L4; diagonal hatching, adult. Data are mean and SD for at least three independent trials, with a minimum of 30 animals from each genotype and stage per trial. srf-3(br6) is a stage-independent Bah control.