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Streptococcus sanguinis  is an early colonizer of the tooth surface and competes with oral pathogens such as  Streptococcus mutans  to maintain oral health. However, little is known about its mechanism of biofilm formation. Here, we show that mutation of the  ciaR  gene, encoding the response regulator of the CiaRH two-component system in  S .  sanguinis  SK36, produced a fragile biofilm. Cell aggregation,  gtfP  gene expression and water-insoluble glucan production were all reduced, which suggested polysaccharide production was decreased in Δ ciaR . RNA sequencing and qRT-PCR revealed that arginine biosynthesis genes ( argR ,  argB ,  argC ,  argG ,  argH  and  argJ ) and two arginine/histidine permease genes (SSA_1568 and SSA_1569) were upregulated in Δ ciaR . In contrast to Δ ciaR , most of strains constructed to contain deletions in each of these genes produced more biofilm and water-insoluble glucan than SK36. A Δ ciaR Δ argB  double mutant was completely restored for the  gtfP  gene expression, glucan production and biofilm formation ability that was lost in Δ ciaR , indicating that  argB  was essential for  ciaR  to regulate biofilm formation. We conclude that by promoting the expression of arginine biosynthetic genes, especially  argB  gene, the  ciaR  mutation reduced polysaccharide production, resulting in the formation of a fragile biofilm in  Streptococcus sanguinis .

ciaR impacts biofilm formation by regulating an arginine biosynthesis pathway in Streptococcus sanguinis SK36