TABLE 1 .

Phenotypic characterization of SagS variants^a

Substitution	Restoration of ΔsagS biofilm-related phenotypes to wild-type levels
	Attachment	Susceptibility to tobramycin
I103	Yes	Yes
D105	Yes	No
A112	Yes	No
S113	Yes	Yes
R116	Intermediate	Yes
R124	Yes	Yes
L126	Yes	No
S127	Yes	No
D128	Yes	No
L130	Yes	Yes
F131	No	Yes
G132b	No	No
K134	Yes	Yes
L141	Yes	Intermediate
G152	Yes	Intermediate
L154	No	Yes
L156b	No	No
D159	Yes	No
D166	Yes	No
F167	Yes	Intermediate
I173	Yes	Intermediate
S177	Yes	Yes
V180	Intermediate	Yes
S182	No	Yes
L183	Yes	No
L185	Intermediate	Yes
L187	Yes	No
L189b	No	Intermediate
V191	Yes	Yes
L197	Yes	Intermediate
T198	Yes	Yes
L204	Yes	No
Q206	Yes	No
L208	No	Yes
D212	Yes	Yes
P213	Yes	No
R218	Yes	Yes
L221	Yes	No
R218	Yes	Intermediate
LILLV	No	Yes
LTKPLVSLIQALb	No	No
IDT	Yes	No
LASASR	Intermediate	Yes
RPLSDFL	Yes	No
ITLLSG	Yes	Intermediate

^aPhenotypic characterization was based on attachment capabilities and susceptibility of biofilms to tobramycin. Wild-type and ΔsagS mutant strains were used as controls. SagS variants with substitutions with underlined residues were chosen for further analyses (see Fig. 4, 5, and 7). Attachment phenotypes correspond to data shown in Fig. 3, while the susceptibility phenotypes are based on data shown in Fig. 6. “Yes” indicates no significant difference from PAO1, “No” indicates significantly different from PAO1, and “Intermediate” indicates significantly different, but values were in between those obtained for PAO1 or the ΔsagS/pMJT-sagS and ΔsagS mutants.

^bThese SagS variants failed to restore both attachment and susceptibility phenotypes by ΔsagS mutant biofilms to wild-type levels, so the respective variants were excluded from subsequent analyses.