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Polymicrobial biofilms play important roles in oral and systemic infections. The oral plaque bacterium  Streptococcus gordonii  is known to attach to the hyphal cell wall of the fungus  Candida albicans  to form corn-cob like structures in biofilms. However, the role of  C. albicans  in formation of polymicrobial biofilms is not completely understood. The objective of this study was to determine the role of  C. albicans  transcription factors in regulation of polymicrobial biofilms and antibiotic tolerance of  S. gordonii . The proteins secreted by  C. albicans  and  S. gordonii  in mixed planktonic cultures were determined using mass spectrometry. Antibiotic tolerance of  S. gordonii  to ampicillin and erythromycin was determined in mixed cultures and mixed biofilms with  C. albicans . Additionally, biofilm formation of  S. gordonii  with  C. albicans  knock-out mutants of 45 transcription factors that affect cell wall integrity, filamentous growth and biofilm formation was determined. Furthermore, these mutants were also screened for antibiotic tolerance in mixed biofilms with  S. gordonii . Analysis of secreted proteomes resulted in the identification of proteins being secreted exclusively in mixed cultures. Antibiotic testing showed that  S. gordonii  had significantly increased survival in mixed planktonic cultures with antibiotics as compared to single cultures.  C. albicans  mutants of transcription factors Sfl2, Brg1, Leu3, Cas5, Cta4, Tec1, Tup1, Rim101 and Efg1 were significantly affected in mixed biofilm formation. Also mixed biofilms of  S. gordonii  with mutants of  C. albicans  transcription factors, Tec1 and Sfl2, had significantly reduced antibiotic tolerance as compared to control cultures. Our data indicates that  C. albicans  may have an important role in mixed biofilm formation as well as antibiotic tolerance of  S. gordonii  in polymicrobial biofilms.  C. albicans  may play a facilitating role than being just an innocent bystander in oral biofilms and infections.

Candida albicanscell wall integrity transcription factors regulate polymicrobial biofilm formation withStreptococcus gordonii