Bacterial transformation: ComFA is a DNA‐dependent ATPase that forms complexes with ComFC and DprA

• Published in: Molecular microbiology Vol. 105; no. 5; pp. 741 - 754
• Main Authors: Diallo, Amy, Foster, Hannah R., Gromek, Katarzyna A., Perry, Thomas N., Dujeancourt, Annick, Krasteva, Petya V., Gubellini, Francesca, Falbel, Tanya G., Burton, Briana M., Fronzes, Rémi
• Format: Journal Article
• Language: English
• Published: England Blackwell Publishing Ltd 01.09.2017; Wiley
• Subjects: Adenosine triphosphatase; Adenosine Triphosphatases; Adenosine Triphosphatases - genetics; Adenosine Triphosphatases - metabolism; Antibiotic resistance; Antibiotics; Bacteria; Bacterial Proteins; Bacterial Proteins - metabolism; Biochemistry, Molecular Biology; Bioinformatics; Biological evolution; Biological Physics; Cellular Biology; Chemical Sciences; Computer Science; Cristallography; Deoxyribonucleic acid; Developmental plasticity; DNA; DNA - metabolism; DNA, Single-Stranded; DNA, Single-Stranded - metabolism; DNA-Binding Proteins; DNA-Binding Proteins - metabolism; Homologous Recombination; Homology; Life Sciences; Membrane Proteins; Membrane Proteins - metabolism; Microbiology; Physics; Protein Binding; Proteins; Rec A Recombinases; Rec A Recombinases - metabolism; Recombination, Genetic; Streptococcus infections; Streptococcus pneumoniae; Streptococcus pneumoniae - genetics; Streptococcus pneumoniae - metabolism; Structural Biology; Transformation, Bacterial; Transformation, Bacterial - genetics; Transformation, Bacterial - physiology
• ISSN: 0950-382X; 1365-2958
• DOI: 10.1111/mmi.13732

Summary Pneumococcal natural transformation contributes to genomic plasticity, antibiotic resistance development and vaccine escape. Streptococcus pneumoniae, like many other naturally transformable species, has evolved sophisticated protein machinery for the binding and uptake of DNA. Two proteins encoded by the comF operon, ComFA and ComFC, are involved in transformation but their exact molecular roles remain unknown. In this study, we provide experimental evidence that ComFA binds to single stranded DNA (ssDNA) and has ssDNA‐dependent ATPase activity. We show that both ComFA and ComFC are essential for the transformation process in pneumococci. Moreover, we show that these proteins interact with each other and with other proteins involved in homologous recombination, such as DprA, thus placing the ComFA‐ComFC duo at the interface between DNA uptake and DNA recombination during transformation. Naturally transformable bacteria use multiprotein machines to import and incorporate DNA. We show that ComFA binds to single stranded DNA (ssDNA), has ssDNA‐stimulated ATPase activity, and that ComFA forms stable complexes with another conserved transformation protein, ComFC. Both ComFA and ComFC are essential for transformation in pneumococci and interact with recombination protein DprA.