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PMID:22328674

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Citation

Carruthers, MD, Tracy, EN, Dickson, AC, Ganser, KB, Munson, RS Jr and Bakaletz, LO (2012) Biological roles of nontypeable Haemophilus influenzae type IV pilus proteins encoded by the pil and com operons. J. Bacteriol. 194:1927-33

Abstract

We previously demonstrated that one or more products of the genes in the pil and com gene clusters of the opportunistic human respiratory pathogen nontypeable Haemophilus influenzae (NTHI) are required for type IV pilus (Tfp) biogenesis and function. Here, we have now demonstrated that the pilABCD and comABCDEF gene clusters are operons and that the product of each gene is essential for normal pilus function. Mutants with nonpolar deletions in each of the 10 pil and com genes had an adherence defect when primary human airway cells were used as the target. These mutants were also diminished in their ability to form a biofilm in vitro and, additionally, were deficient in natural transformation. Collectively, our data demonstrate that the product of each gene within these operons is required for the normal biogenesis and/or function of NTHI Tfp. Based on the similarity of PilA to other type IV pilins, we further predicted that the product of the pilA gene would be the major pilin subunit. Toward that end, we also demonstrated by immunogold labeling and mass spectrometry that PilA is indeed the majority type IV pilin protein expressed by NTHI. These new observations set the stage for experiments designed to dissect the function of each of the proteins encoded by genes within the pil and com gene clusters. The ability to characterize individual proteins with vital roles in NTHI colonization or pathogenesis has the potential to reduce the burden of NTHI-induced diseases through development of a Tfp-derived vaccine or a pilus-directed therapeutic.

Links

PubMed PMC3318474 Online version:10.1128/JB.06540-11

Keywords

Bacterial Adhesion; Bacterial Proteins/genetics; Bacterial Proteins/metabolism; Biofilms; Cells, Cultured; Epithelial Cells/microbiology; Fimbriae, Bacterial/genetics; Fimbriae, Bacterial/metabolism; Gene Expression Regulation, Bacterial/physiology; Haemophilus influenzae/genetics; Haemophilus influenzae/metabolism; Humans; Multigene Family; Mutation; Operon; Respiratory Mucosa/cytology

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

HAEI8:Q4QNP4

GO:1900231: regulation of single-species biofilm formation on inanimate substrate

ECO:0000315:

P

Figure 2A&B. NTHI pilA mutants had significantly reduced biofilm thickness and biomass compared to the wildtype.

complete
CACAO 5958

HAEI8:Q4QNP4

GO:0052001: Type IV pili-dependent localized adherence to host

ECO:0000315:

P

Figure 3. NTHI pilA mutants had significant adherence defects on normal human bronchial epithelial cells compared to the wildtype.

complete
CACAO 5959

HAEI8:Q4QNP4

involved_in

GO:0052001: type IV pili-dependent localized adherence to host

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

HAEI8:Q4QNP4

involved_in

GO:1900231: regulation of single-species biofilm formation on inanimate substrate

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

HAEI8:Q4QNP5

GO:1900231: regulation of single-species biofilm formation on inanimate substrate

ECO:0000315:

P

Figure 2A&B. NTHI pilB mutants had significantly reduced biofilm thickness and biomass compared to the wildtype.

complete
CACAO 5960

HAEI8:Q4QNP5

GO:0052001: Type IV pili-dependent localized adherence to host

ECO:0000315:

P

Figure 3. NTHI pilB mutants had significant adherence defects on normal human bronchial epithelial cells compared to the wildtype.

complete
CACAO 5961

HAEI8:Q4QNP5

involved_in

GO:0052001: type IV pili-dependent localized adherence to host

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

HAEI8:Q4QNP5

involved_in

GO:1900231: regulation of single-species biofilm formation on inanimate substrate

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

HAEI8:Q4QNP6

GO:1900231: regulation of single-species biofilm formation on inanimate substrate

ECO:0000315:

P

Figure 2A&B. NTHI pilC mutants had significantly reduced biofilm thickness and biomass compared to the wildtype.

complete
CACAO 5962

HAEI8:Q4QNP6

GO:0052001: Type IV pili-dependent localized adherence to host

ECO:0000315:

P

Figure 3. NTHI pilC mutants had significant adherence defects on normal human bronchial epithelial cells compared to the wildtype.

complete
CACAO 5963

HAEI8:Q4QNP6

involved_in

GO:0052001: type IV pili-dependent localized adherence to host

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

HAEI8:Q4QNP6

involved_in

GO:1900231: regulation of single-species biofilm formation on inanimate substrate

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

HAEI8:Q4QNP7

GO:1900231: regulation of single-species biofilm formation on inanimate substrate

ECO:0000315:

P

Figure 2A&B. NTHI pilD mutants had significantly reduced biofilm thickness and biomass compared to the wildtype.

complete
CACAO 5964

HAEI8:Q4QNP7

GO:0052001: Type IV pili-dependent localized adherence to host

ECO:0000315:

P

Figure 3. NTHI pilD mutants had significant adherence defects on normal human bronchial epithelial cells compared to the wildtype.

complete
CACAO 5965

HAEI8:Q4QNP7

involved_in

GO:0052001: type IV pili-dependent localized adherence to host

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

HAEI8:Q4QNP7

involved_in

GO:1900231: regulation of single-species biofilm formation on inanimate substrate

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

HAEI8:Q4QNA6

GO:0052001: Type IV pili-dependent localized adherence to host

ECO:0000315:

P

Figure 3. NTHI comA mutants had significant adherence defects on normal human bronchial epithelial cells compared to the wildtype.

complete
CACAO 5967

HAEI8:Q4QNA6

involved_in

GO:0052001: type IV pili-dependent localized adherence to host

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

HAEI8:Q4QNA8

GO:1900231: regulation of single-species biofilm formation on inanimate substrate

ECO:0000315:

P

Figure 2A&B. NTHI comC mutants had significantly reduced biofilm thickness and biomass compared to the wildtype.

complete
CACAO 5968

HAEI8:Q4QNA8

GO:0052001: Type IV pili-dependent localized adherence to host

ECO:0000315:

P

Figure 3. NTHI comC mutants had significant adherence defects on normal human bronchial epithelial cells compared to the wildtype.

complete
CACAO 5969

HAEI8:Q4QNA8

involved_in

GO:0052001: type IV pili-dependent localized adherence to host

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

HAEI8:Q4QNA8

involved_in

GO:1900231: regulation of single-species biofilm formation on inanimate substrate

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

HAEI8:Q4QNA9

GO:0052001: Type IV pili-dependent localized adherence to host

ECO:0000315:

P

Figure 3. NTHI comD mutants had significant adherence defects on normal human bronchial epithelial cells compared to the wildtype.

complete
CACAO 5970

HAEI8:Q4QNA9

involved_in

GO:0052001: type IV pili-dependent localized adherence to host

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

HAEI8:Q4QNB0

GO:0052001: Type IV pili-dependent localized adherence to host

ECO:0000315:

P

Figure 3. NTHI comE mutants had significant adherence defects on normal human bronchial epithelial cells compared to the wildtype.

complete
CACAO 5971

HAEI8:Q4QNB0

involved_in

GO:0052001: type IV pili-dependent localized adherence to host

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

HAEI8:Q4QNB1

GO:0042710: biofilm formation

ECO:0000315:

P

Figure 3. NTHI comF mutants had significant adherence defects on normal human bronchial epithelial cells compared to the wildtype.

complete
CACAO 5972


See also

References

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