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Identification of PblB mediating galactose-specific adhesion in a successful Streptococcus pneumoniae clone.

Hsieh YC, Lin TL, Lin CM, Wang JT - Sci Rep (2015)

Bottom Line: Preincubation of NTUH-P15 with D-galactose resulted in decreases of adherence to A549 cell in a dose-dependent manner.Challenge of mice with NTUH-P15, isogenic pblB mutant and pblB complementation strains determined that PblB was required for bacterial persistence in the nasopharynx and lung.PblB, as an adhesin mediating the galactose-specific adhesion activity of pneumococci, promote pneumococcal clonal success.

View Article: PubMed Central - PubMed

Affiliation: Department of Pediatrics, Chang Gung Children's Hospital, Chang Gung Memorial Hospital, Chang Gung University, College of Medicine, Taoyuan, Taiwan.

ABSTRACT
The pneumococcal genome is variable and there are minimal data on the influence of the accessory genome on phenotype. Pneumococcal serotype 14 sequence type (ST) 46 had been the most prevalent clone causing pneumonia in children in Taiwan. A microarray was constructed using the genomic DNA of a clinical strain (NTUH-P15) of serotype 14 ST46. Using DNA hybridization, genomic variations in NTUH-P15 were compared to those of 3 control strains. Microarray analysis identified 7 genomic regions that had significant increases in hybridization signals in the NTUH-P15 strain compared to control strains. One of these regions encoded PblB, a phage-encoded virulence factor implicated (in Streptococcus mitis) in infective endocarditis. The isogenic pblB mutant decreased adherence to A549 human lung epithelial cell compared to wild-type NTUH-P15 strain (P = 0.01). Complementation with pblB restored the adherence. PblB is predicted to contain a galactose-binding domain-like region. Preincubation of NTUH-P15 with D-galactose resulted in decreases of adherence to A549 cell in a dose-dependent manner. Challenge of mice with NTUH-P15, isogenic pblB mutant and pblB complementation strains determined that PblB was required for bacterial persistence in the nasopharynx and lung. PblB, as an adhesin mediating the galactose-specific adhesion activity of pneumococci, promote pneumococcal clonal success.

No MeSH data available.


Related in: MedlinePlus

Schematic representation of PblB of Streptococcus pneumoniae.A. The host specificity protein gene and pblB gene (black squares) residing within a 33-kb temperate bacteriophage located in an insertion site between the purA (adenylosuccinate synthetase) and tadA (tRNA-specific adenosine deaminase) (gray squares) genes in Streptococcus pneumoniae P1031. B. Analysis by using the SMART program (http://smart.embl-heidelberg.de/). PblB of Streptococcus pneumoniae NTUH-P15 is composed of a signal peptide, a coiled-coil region, four internal repeats, and a galactose-binding domain-like region. PblB of Streptococcus mitis SF100 is composed of a coiled-coil region and a peptidase domain. C. Analysis by using BLAST. PblB of Streptococcus pneumoniae NTUH-P15 is composed of a prophage endopeptidase tail and a carbonhydrate binding domain. PblB of Streptococcus mitis SF100 is composed of a prophage tail and a peptidase domain.
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f2: Schematic representation of PblB of Streptococcus pneumoniae.A. The host specificity protein gene and pblB gene (black squares) residing within a 33-kb temperate bacteriophage located in an insertion site between the purA (adenylosuccinate synthetase) and tadA (tRNA-specific adenosine deaminase) (gray squares) genes in Streptococcus pneumoniae P1031. B. Analysis by using the SMART program (http://smart.embl-heidelberg.de/). PblB of Streptococcus pneumoniae NTUH-P15 is composed of a signal peptide, a coiled-coil region, four internal repeats, and a galactose-binding domain-like region. PblB of Streptococcus mitis SF100 is composed of a coiled-coil region and a peptidase domain. C. Analysis by using BLAST. PblB of Streptococcus pneumoniae NTUH-P15 is composed of a prophage endopeptidase tail and a carbonhydrate binding domain. PblB of Streptococcus mitis SF100 is composed of a prophage tail and a peptidase domain.

Mentions: Analysis of the complete genome of the P1031 strain suggested that the host specific protein gene and pblB gene reside within a 33-kb temperate bacteriophage located in an insertion site between the purA (adenylosuccinate synthetase) and tadA (tRNA-specific adenosine deaminase) genes (Fig. 2A). We cloned and sequenced the corresponding chromosomal region (i.e., flanking the host specific protein gene and pblB gene) from NTUH-P15. A total of 7.1 kb, including 0.7 kb upstream and 0.5 kb downstream, was sequenced. Analysis of sequence suggested that the host specific protein gene and pblB gene of NTUH-P15 would be transcribed together, in contrast to the separate transcription predicted for the loci in S. pneumoniae P1031. PblB of NTUH-P15 (GenBank accession number AB679266) is predicted to encode a 213-kDa protein with a pI of 8.98. By Pairwise Sequence Alignment (www.ebi.ac.uk/Tools/psa/), the predicted PblB of NTUH-P15 shared 24.3% sequence identity and 40.3% similarity with PblB in S. mitis. On the basis of its amino acid sequence, PblB is predicted to form a signal peptide, a coiled-coil region, 4 internal repeats, and a galactose-binding domain-like region located at the carboxy terminus by the SMART program (http://smart.embl-heidelberg.de/) (Fig. 2B). By BLAST analysis, PblB is predicted to form a prophage endopeptidase tail and a carbonhydrate binding domain (Fig. 2C).


Identification of PblB mediating galactose-specific adhesion in a successful Streptococcus pneumoniae clone.

Hsieh YC, Lin TL, Lin CM, Wang JT - Sci Rep (2015)

Schematic representation of PblB of Streptococcus pneumoniae.A. The host specificity protein gene and pblB gene (black squares) residing within a 33-kb temperate bacteriophage located in an insertion site between the purA (adenylosuccinate synthetase) and tadA (tRNA-specific adenosine deaminase) (gray squares) genes in Streptococcus pneumoniae P1031. B. Analysis by using the SMART program (http://smart.embl-heidelberg.de/). PblB of Streptococcus pneumoniae NTUH-P15 is composed of a signal peptide, a coiled-coil region, four internal repeats, and a galactose-binding domain-like region. PblB of Streptococcus mitis SF100 is composed of a coiled-coil region and a peptidase domain. C. Analysis by using BLAST. PblB of Streptococcus pneumoniae NTUH-P15 is composed of a prophage endopeptidase tail and a carbonhydrate binding domain. PblB of Streptococcus mitis SF100 is composed of a prophage tail and a peptidase domain.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4508584&req=5

f2: Schematic representation of PblB of Streptococcus pneumoniae.A. The host specificity protein gene and pblB gene (black squares) residing within a 33-kb temperate bacteriophage located in an insertion site between the purA (adenylosuccinate synthetase) and tadA (tRNA-specific adenosine deaminase) (gray squares) genes in Streptococcus pneumoniae P1031. B. Analysis by using the SMART program (http://smart.embl-heidelberg.de/). PblB of Streptococcus pneumoniae NTUH-P15 is composed of a signal peptide, a coiled-coil region, four internal repeats, and a galactose-binding domain-like region. PblB of Streptococcus mitis SF100 is composed of a coiled-coil region and a peptidase domain. C. Analysis by using BLAST. PblB of Streptococcus pneumoniae NTUH-P15 is composed of a prophage endopeptidase tail and a carbonhydrate binding domain. PblB of Streptococcus mitis SF100 is composed of a prophage tail and a peptidase domain.
Mentions: Analysis of the complete genome of the P1031 strain suggested that the host specific protein gene and pblB gene reside within a 33-kb temperate bacteriophage located in an insertion site between the purA (adenylosuccinate synthetase) and tadA (tRNA-specific adenosine deaminase) genes (Fig. 2A). We cloned and sequenced the corresponding chromosomal region (i.e., flanking the host specific protein gene and pblB gene) from NTUH-P15. A total of 7.1 kb, including 0.7 kb upstream and 0.5 kb downstream, was sequenced. Analysis of sequence suggested that the host specific protein gene and pblB gene of NTUH-P15 would be transcribed together, in contrast to the separate transcription predicted for the loci in S. pneumoniae P1031. PblB of NTUH-P15 (GenBank accession number AB679266) is predicted to encode a 213-kDa protein with a pI of 8.98. By Pairwise Sequence Alignment (www.ebi.ac.uk/Tools/psa/), the predicted PblB of NTUH-P15 shared 24.3% sequence identity and 40.3% similarity with PblB in S. mitis. On the basis of its amino acid sequence, PblB is predicted to form a signal peptide, a coiled-coil region, 4 internal repeats, and a galactose-binding domain-like region located at the carboxy terminus by the SMART program (http://smart.embl-heidelberg.de/) (Fig. 2B). By BLAST analysis, PblB is predicted to form a prophage endopeptidase tail and a carbonhydrate binding domain (Fig. 2C).

Bottom Line: Preincubation of NTUH-P15 with D-galactose resulted in decreases of adherence to A549 cell in a dose-dependent manner.Challenge of mice with NTUH-P15, isogenic pblB mutant and pblB complementation strains determined that PblB was required for bacterial persistence in the nasopharynx and lung.PblB, as an adhesin mediating the galactose-specific adhesion activity of pneumococci, promote pneumococcal clonal success.

View Article: PubMed Central - PubMed

Affiliation: Department of Pediatrics, Chang Gung Children's Hospital, Chang Gung Memorial Hospital, Chang Gung University, College of Medicine, Taoyuan, Taiwan.

ABSTRACT
The pneumococcal genome is variable and there are minimal data on the influence of the accessory genome on phenotype. Pneumococcal serotype 14 sequence type (ST) 46 had been the most prevalent clone causing pneumonia in children in Taiwan. A microarray was constructed using the genomic DNA of a clinical strain (NTUH-P15) of serotype 14 ST46. Using DNA hybridization, genomic variations in NTUH-P15 were compared to those of 3 control strains. Microarray analysis identified 7 genomic regions that had significant increases in hybridization signals in the NTUH-P15 strain compared to control strains. One of these regions encoded PblB, a phage-encoded virulence factor implicated (in Streptococcus mitis) in infective endocarditis. The isogenic pblB mutant decreased adherence to A549 human lung epithelial cell compared to wild-type NTUH-P15 strain (P = 0.01). Complementation with pblB restored the adherence. PblB is predicted to contain a galactose-binding domain-like region. Preincubation of NTUH-P15 with D-galactose resulted in decreases of adherence to A549 cell in a dose-dependent manner. Challenge of mice with NTUH-P15, isogenic pblB mutant and pblB complementation strains determined that PblB was required for bacterial persistence in the nasopharynx and lung. PblB, as an adhesin mediating the galactose-specific adhesion activity of pneumococci, promote pneumococcal clonal success.

No MeSH data available.


Related in: MedlinePlus