Involvement of the adc operon and manganese homeostasis in Streptococcus gordonii biofilm formation.
Pioneer oral bacteria, including Streptococcus gordonii, initiate the formation of oral biofilms on tooth surfaces, which requires differential expression of genes that recognize unique environmental cues. An S. gordonii::Tn917-lac biofilm-defective mutant was isolated by using an in vitro biofilm formation assay. Subsequent inverse PCR and sequence analyses identified the transposon insertion to be near the 3' end of an open reading frame (ORF) encoding a protein homologous to a Streptococcus pneumoniae repressor, AdcR. The S. gordonii adc operon, consisting of the four ORFs adcR, adcC, adcB, and adcA, is homologous to the adc operon of S. pneumoniae, which plays a role in zinc and/or manganese transport and genetic competence in S. pneumoniae. AdcR is a metal-dependent repressor protein containing a putative metal-binding site, AdcC contains a consensus-binding site for ATP, AdcB is a hydrophobic protein with seven hydrophobic membrane-spanning regions, and AdcA is a lipoprotein permease with a putative metal-binding site. The three proteins (AdcC through -A) are similar to those of the binding-lipoprotein-dependent transport system of gram-positive bacteria. Reverse transcriptase PCR confirmed that adcRCBA are cotranscribed as an operon in S. gordonii and that the transposon insertion in S. gordonii adcR::Tn917-lac had resulted in a polar mutation. Expression of adcR, measured by the beta-galactosidase activity of the adcR::Tn917-lac mutant, was growth phase dependent and increased when the mutant was grown in media with high levels of manganese (>1 mM) and to a lesser extent in media with zinc, indicating that AdcR may be a regulator at high levels of extracellular manganese. A nonpolar inactivation of adcR generated by allelic replacement resulted in a biofilm- and competence-defective phenotype. The biofilm-defective phenotype observed suggests that AdcR is an active repressor when synthesized and acts at a distant site(s) on the chromosome. Thus, the adc operon is involved in manganese acquisition in S. gordonii and manganese homeostasis and appears to modulate sessile growth in this bacterium.
Loo
C Y
CY
Department of Pediatric Dentistry, Goldman School of Dental Medicine, Boston University, Boston, Massachusetts 02118, USA.
Mitrakul
K
K
Voss
I B
IB
Hughes
C V
CV
Ganeshkumar
N
N
D018528:ATP-Binding Cassette Transporters / Q000235:genetics* / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics; D018441:Biofilms / Q000254:growth & development*; D001692:Biological Transport; D003470:Culture Media; D004251:DNA Transposable Elements; D015964:Gene Expression Regulation, Bacterial; D006706:Homeostasis; D008345:Manganese / Q000378:metabolism*; D008565:Membrane Proteins / Q000235:genetics; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D016366:Open Reading Frames; D009876:Operon / Q000502:physiology*; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D013291:Streptococcus / Q000235:genetics / Q000502:physiology*; D015032:Zinc / Q000378:metabolism
D018528:ATP-Binding Cassette Transporters; C114936:AdcA protein, Streptococcus; C114935:AdcB protein, Streptococcus; C114934:AdcC protein, Streptococcus; D001426:Bacterial Proteins; D003470:Culture Media; D004251:DNA Transposable Elements; D008565:Membrane Proteins; D012097:Repressor Proteins; D008345:Manganese; D000255:Adenosine Triphosphate; D015032:Zinc