Fig 1.
Annotated motility cluster in A. fabrum.
Shown in gray are genes that were not disrupted in the screen. Shown in white are genes that were disrupted in the screen and are of strongly suspected function. Shown in black are genes that were disrupted in the screen and were subjected to further analysis. White dots on top of the genes show that a transposon landed in rightward orientation (referring to kanR transcription), while black dots indicate that a transposon landed in leftward orientation. A 16-kb region not shown in the figure includes genes that are not predicted to be involved in flagellar assembly, and no genes in this region were disrupted in the screen.
Fig 2.
The putative structure of the A. fabrum flagellum and the proteins from which it is constituted.
The diagram includes the orientation of the flagellum with respect to major cell envelope components: Outer membrane (OM), peptidoglycan (PG), and the inner membrane (IM). Also labeled are the main functional units of the flagellum and the specific proteins from which they are made, all of which were implicated in this genetic screen. Proteins with regulatory or unknown functions are listed in the box.
Fig 3.
Synteny of four gene cluster required for motility in Sinorhizobium meliloti and Agrobacterium fabrum.
ATU0585 was renamed flgN and ATU8132 was renamed motF.
Fig 4.
In-frame deletions and complementation experiments of the four genes subjected to this study.
Motility assay for strains with each in-frame deletion and their corresponding complementation assays. Swim rings were imaged 48 h after inoculation. Shown are the averages of swim ring diameter for four replicates per strain in millimeters (mm) and standard deviation from the mean. Statistical analysis is shown in S3 and S4 Figs. Strategy used for in-frame deletions is depicted in S5A Fig.
Fig 5.
ΔmotF has a suppressible phenotype.
Strains were inoculated and allowed to swim for 3 (top) or 6 (bottom) days. The ΔflaF strain serves as a non-suppressible control. Below each strain is the average of swim ring diameters in millimeters (mm) for four replicates per strain and the standard deviation from the mean. Statistical analysis is shown in S7 Fig.
Fig 6.
Characterization of the ΔvisNR deletion strain.
(A) Complementation test in which ΔvisNR strains harbor plasmids indicated in S1 Fig. Shown are the averages of swim ring diameters in millimeters (mm) in four replicates per strain and standard deviation from the mean. Statistical analysis is shown in S9 Fig. (B) The left image shows a plate with ΔvisNR colonies, and the right image shows a plate with parent strain BB01. Both strains were grown under the same conditions and images are shown at the same scale.