Article Figures & Data
Figures
- Fig 1
Growth of unmarked H. influenzae CRP-S mutants in rich medium. Cells were grown in sBHI broth in a BioScreen incubator with the OD600 recorded every 10 min. Each line shows the mean of 8 to 12 replicates for one strain. Optical densities lower than 0.02 are not shown. To account for differences in inoculum sizes, the lines have been shifted along the x axis to superimpose the growth curves of all mutants. Positive and negative controls are shown as black lines (full lines, wild type; dashed lines, sxy mutant).
- Fig 2
Competence phenotypes of unmarked H. influenzae CRP-S mutants. Each point represents the mean of at least three biological replicates. Mean values for positive (wild-type) and negative (sxy mutant) controls are shown as an open circle and an open square, respectively. Mutants in which competence is abolished are shaded.
- Fig 3
DNA uptake and transformation by H. influenzae CRP-S mutants. (A) DNA uptake by unmarked mutants. (B) Operon structure of the CRP-S genes of H. influenzae. (C) Transformation frequencies of unmarked mutants. (D) Transformation frequencies of marked mutants. In panels A, C, and D, vertical bars represent the means of at least three biological replicates ± standard deviations. The range of wild-type and sxy mutant control values are shown as gray horizontal bars. Statistically significant differences from the wild type (P value of <0.05, determined by a two-tailed t test) are indicated with asterisks.
- Fig 4
Competence phenotypes of the H. influenzae comI and comJ mutants. The operon structure is shown at the top: the comI and comJ genes are divergently transcribed from the comABCDEF operon and its CRP-S promoter. Triangles show the locations of previously described transposon insertions (100, 101). The uptake and transformation phenotypes of the comI and comJ mutants that we created (I-, unmarked comI; J-, unmarked comJ; Isp, marked comI::Spec; Jsp, marked comJ::Spec) and of previously described transposon mutants (100, 101) are shown at the bottom. Each bar represents the mean of at least three biological replicates ± the standard deviation.
Tables
- Table 1
Genes and proteins of the CRP-S regulon of H. influenzaea
Gene Locus tag Reported H. influenzae mutant phenotype (reference[s]) H. influenzae protein sequence predictionc Homolog(s) Function(s) of homolog(s) (reference[s]) pilA HI0299 Uptake−, TF− (32, 104) 16 kDa, pI 9.3; prepilin peptidase signal; periplasmic and extracellular locations pilE (Neisseria), pilA (Pseudomonas) Major pilin subunit (2, 58, 73, 77) pilB HI0298 Uptake−, TF− (32) 53 kDa, pI 5.7; no signal detected; cytoplasmic location pilB (Neisseria, Pseudomonas) ATP-dependent extension of pilus subunits (39, 74) pilC HI0297 Uptake−, TF− (32) 47 kDa, pI 9.4; no signal detected; IM location, 3 transmembrane helices pilC (Neisseria, Pseudomonas) Pilus assembly (39, 74, 102) pilD HI0296 None available 27 kDa, pI 9.3; no signal detected; IM location, 7 transmembrane helices pilD (Neisseria, Pseudomonas) Prepilin peptidase (25, 39, 74, 75) comA HI0439 Uptake−, TF− (101) 32 kDa, pI 5.7; no signal detected; IM location pilM (Neisseria, Pseudomonas) Pilus assembly (7, 18, 65) comB HI0438 None available 20 kDa, pI 8.3; noncleavable signal peptide; location unknown pilN (Neisseria, Pseudomonas) Pilus assembly (7, 18, 65, 86, 98) comC HI0437 Uptake−, TF− (101) 20 kDa, pI 9.3; signal peptidase I signal; location unknown pilO (Neisseria, Pseudomonas) Pilus assembly (7, 18, 65, 86, 98) comD HI0436 None available 16 kDa, pI 4.6; signal peptidase II signal; location unknown pilP (Neisseria, Pseudomonas) Pilus assembly, IM lipoprotein; interacts with secretin (7, 8, 18, 65, 98) comE HI0435 Uptake−, TF− (32, 101) 49 kDa, pI 7.4; signal peptidase I signal; OM location pilQ (Neisseria, Pseudomonas) OM secretin, binds T4P and DNA (5, 28, 65) comF HI0434 WT uptake, TF− (100, 101) 27 kDa, pI 10.3; no signal detected; cytoplasmic location comF (Neisseria, Pseudomonas) Function unknown comN HI0938 None available 20 kDa, pI 9.6; prepilin peptidase signal; location unknown No homolog found Function unknown comO HI0939 TF− (107) 27 kDa, pI 9.6; signal peptidase I signal; location unknown No homolog found Function unknown comP HI0940 None available 25 kDa, pI 9; signal peptidase I signal; location unknown No homolog found Function unknown comQ HI0941 None available 12 kDa, pI 9.9; signal peptidase I signal; location unknown No homolog found Function unknown comE1 HI008 None available 12 kDa, pI 8.3; signal peptidase I signal; location unknown comE (Neisseria, Pseudomonas), comEA (Bacillus) DNA binding protein (23, 45, 72, 82) HI0365 HI0365 WT uptake and TF (107) 44 kDa, pI 6.7; no signal detected; cytoplasmic location; rRNA large subunit methyltransferase N; Fe-S cluster redox enzyme Hypothetical protein (Neisseria, Pseudomonas) Function unknown pilF2 HI0366 Uptake−, TF− (107) 21 kDa, pI 8.9; signal peptidase I signal; location unknown; tetratricopeptide repeats pilW (Neisseria), pilF (Pseudomonas) Pilotin, stabilizes secretin multimers (3, 19, 50, 51, 95, 105) rec2 HI0061 WT uptake, TF− (68, 90) 90 kDa, pI 9.7; no signal detected; IM location, 13 transmembrane helices comA (Neisseria), rec2 (Pseudomonas), comEC (Bacillus) Putative DNA membrane channel (34) ligA HI1182, HI1183b WT uptake, TF down (107) 31 kDa, pI 9.2; signal peptidase I signal; location unknown adl (Neisseria) Periplasmic ATP-dependent DNA ligase (63) HI0659 HI0659 None available 11 kDa, pI 9.2; no signal detected; cytoplasmic location; HTH motif No homolog found Function unknown HI0660 HI0660 None available 14 kDa, pI 9.46; no signal detected; cytoplasmic location; putative Holliday junction resolvase No homolog found Function unknown HI1631 HI1631 None available 22 kDa, pI 9.5; no signal detected; location unknown; restriction endonuclease-like superfamily; putative Holliday junction resolvase No homolog found Function unknown comM HI1117 WT uptake, TF down (42) 56 kDa, pI 9.2; no signal detected; cytoplasmic location; putative Mg chelatase subunit comM (Neisseria, Pseudomonas) Function unknown dprA HI0985 TF− (48, 100) 42 kDa, pI 6.3; no signal detected; cytoplasmic location dprA (Neisseria), smf (Pseudomonas) Protects DNA from degradation in cytoplasm (4) radC HI0952 None available 25 kDa, pI 6.2; no signal detected; cytoplasmic location radC (Pseudomonas) Replication fork stabilization and repair (6, 87) ssb HI0250 None available 19 kDa, pI 5.3; no signal detected; cytoplasmic location ssb (Neisseria, Pseudomonas) Ubiquitous single-stranded-DNA binding protein (83)
