Comparative Analysis of Cellulophaga algicola and Flavobacterium johnsoniae Gliding Motility

Abstract

Gliding motility is common in members of the phylum Bacteroidetes, including Flavobacterium johnsoniae and Cellulophaga algicola. F. johnsoniae gliding has been extensively studied and involves rapid movement of the cell surface adhesin SprB. Genetic analysis of C. algicola allowed a comparative analysis of gliding. Sixty-three HimarEm1-induced mutants that formed nonspreading colonies were characterized. Each had an insertion in an ortholog of an F. johnsoniae motility gene, highlighting similarities between the motility systems. Differences were also observed. C. algicola lacks orthologs of the F. johnsoniae motility genes gldA, gldF, and gldG that are thought to encode the components of an ATP-binding cassette (ABC) transporter. In addition, mutations in any of 12 F. johnsoniae gld genes result in complete loss of motility, whereas all C. algicola gld mutants retained slight residual motility. This may indicate that C. algicola has multiple motility systems, that the motility proteins exhibit partial redundancy of function, or that essential components of the motility machinery of both C. algicola and F. johnsoniae remain to be discovered. IMPORTANCEThe development of genetic tools for C. algicola and comparative analysis of F. johnsoniae and C. algicola motility mutants identified similarities and differences between their gliding motility machineries. Gliding motility is common in the phylum Bacteroidetes. Proteins that are important for gliding in both C. algicola and F. johnsoniae are potential core components of the Bacteroidetes gliding motility machinery. Many bacteria belonging to the phylum Bacteroidetes exhibit rapid gliding motility over surfaces (1). Gliding has been most well studied in Flavobacterium johnsoniae, in which the disruption of any of 11 gld genes (gldA, gldB, gldD, gldF, gldG, gldH, gldI, gldJ, gldK, gldL, and gldM) or deletion of gldN and its paralog gldO results in a complete loss of motility (2-10). These gld mutants form nonspreading colonies on agar, and individual cells exhibit no movement on glass or agar. Mutations in other F. johnsoniae genes (sprA, sprB, sprC, sprD, sprE, sprF, and sprT) result in partial motility defects (11-15). These spr mutants form nonspreading colonies composed of cells that exhibit some limited ability to glide on glass surfaces.sprB encodes an adhesin that is propelled rapidly along the cell surface and is thought to account for much of the gliding movement of F. johnsoniae (11, 16). The F. johnsoniae genome encodes other SprB-like proteins, such as RemA, which explains why sprB mutants exhibit some residual motility (17). Seven proteins (GldK, GldL, GldM, GldN, SprA, SprE, and SprT) comprise the type IX secretion system (T9SS) that is required for the delivery of SprB to the cell surface and for the secretion of dozens of other cell surface and extracellular proteins (14,15,18,19). SprF is also needed for the secretion of SprB but not for the secretion of other proteins targeted to the T9SS (13). The motor that propels SprB along the cel...

Genetic analyses unravel the crucial role of a horizontally acquired alginate lyase for brown algal biomass degradation by Zobellia galactanivorans

“…Techniques to construct gene deletions and other unmarked site‐directed mutations in the chromosomes of several members of the Bacteroidetes have been developed (Pumbwe et al ., ; Koropatkin et al ., ; Rhodes et al ., ; Wang et al ., ; Zhu and McBride, ; Li et al ., ; Zhu and McBride, ). Most of these cannot be used on wild type cells but instead require strains that already have specific chromosomal mutations to allow counterselection.…”

“…To test this, we determined the ability of pYT313 to facilitate targeted gene deletion in wild type cells of three distantly related members of the phylum Bacteroidetes . Markerless in‐frame deletions of Cytophaga hutchinsonii endoglucanase‐encoding gene cel5A (Zhu et al ., ), F. johnsoniae motility gene sprF (Rhodes et al ., ) and Cellulophaga algicola motility gene gldB (Zhu and McBride, ) were obtained (Supporting Information Fig. S7 and Fig.…”

“…Zobellia galactanivorans strains were grown at 30°C in Cytophaga medium (DSMZ medium 172) which consisted of 1.0 g L −1 yeast extract, 1.0 g L −1 tryptone, 24.7 g L −1 NaCl, 0.7 g L −1 KCl, 6.3 g L −1 MgSO 4 ·7H 2 O, 4.6 g L −1 MgCl 2 ·6H 2 O, 1.2 g L −1 CaCl 2 ·2H 2 O, 0.2 g L −1 NaHCO 3 , pH 7.2. Marine conjugation medium (Zhu and McBride, ) consisted of 1.0 g L −1 yeast extract, 1.0 g L −1 tryptone, 5 g L −1 NaCl, 0.35 g L −1 KCl, 3.15 g L −1 MgSO 4 ·7H 2 O, 2.3 g L −1 MgCl 2 ·6H 2 O, 0.6 g L −1 CaCl 2 ·2H 2 O, 0.1 g L −1 NaHCO 3 (pH 7.2). Marine conjugation agar was identical except for the addition of 15 g agar L −1 .…”

DivIVA Controls Progeny Morphology and Diverse ParA Proteins Regulate Cell Division or Gliding Motility in Bdellovibrio bacteriovorus

“…Bacterial gliding is a process characterized by the nonflagellar movement of a single cell on a solid surface and has been previously observed in many bacterial species, including B. bacteriovorus (Spormann, 1999;Mendez et al, 2008, Lambert et al, 2011Asada et al, 2012, Zhu andMcBride, 2016). Gliding motility can be subdivided into two categories; social (S)motility, surface movement using pilus retraction in Myxococcus xanthus; and adventurous (A)-gliding motility, characterized by the movement of individual cells (McBride and Zhu, 2013;Jakobczak et al, 2015, Zhu andMcBride, 2016). B. bacteriovorus is known to exhibit a form of A-motility which uses gliding engines and is independent of pili (Lambert et al, 2011).…”

Comparative genomic analysis of Flavobacteriaceae: insights into carbohydrate metabolism, gliding motility and secondary metabolite biosynthesis.

“…In this study, we followed the definition of flavobacterial clades as proposed by McBride actively gliding even though their genomes were lacking the genes encoding this ABC transporter (21). This might suggest either the involvement of another ABC transporter, common in most organisms, or a non-essential role of this ABC transporter in Bacteroidetes gliding motility (21,23,67). The T9SS exports the motility adhesins RemA and SprB which are propelled along the cell surface by the gliding motor, resulting in cell movement of F. johnsoniae (21,25,68).…”