title: The secretion ATPase ComGA is required for the binding and transport of transforming DNA.
abstract: Transformation requires specialized proteins to facilitate the binding and uptake of DNA. The genes of the Bacillus subtilis comG operon (comGA-G) are required for transformation and to assemble a structure, the pseudopilus, in the cell envelope. No role for the pseudopilus has been established and the functions of the individual comG genes are unknown. We show that among the comG genes, only comGA is absolutely required for DNA binding to the cell surface. ComEA, an integral membrane DNA-binding protein plays a minor role in the initial binding step, while an unidentified protein which communicates with ComGA must be directly responsible for binding to the cell. We show that the use of resistance to DNase to measure 'DNA uptake' reflects the movement of transforming DNA to a protected state in which it is not irreversibly associated with the protoplast, and presumably resides outside the cell membrane, in the periplasm or associated with the cell wall. We suggest that ComGA is needed for the acquisition of DNase resistance as well as for the binding of DNA to the cell surface. Finally, we show that the pseudopilus is required for DNA uptake and we offer a revised model for the transformation process.
authors.affiliation: Public Health Research Institute Center, New Jersey Medical School, University of Medicine and Dentistry of New Jersey, Newark, NJ 07103, USA.
authors.forename: Kenneth
authors.initials: K
authors.lastname: Briley
authors.forename: Angella
authors.initials: A
authors.lastname: Dorsey-Oresto
authors.forename: Peter
authors.initials: P
authors.lastname: Prepiak
authors.forename: Miguel J
authors.initials: MJ
authors.lastname: Dias
authors.forename: Jessica M
authors.initials: JM
authors.lastname: Mann
authors.forename: David
authors.initials: D
authors.lastname: Dubnau
mesh_terms: D000251:Adenosine Triphosphatases / Q000378:metabolism*; D000595:Amino Acid Sequence; D001412:Bacillus subtilis / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism*; D004269:DNA, Bacterial / Q000235:genetics / Q000378:metabolism*; D003851:Deoxyribonucleases / Q000378:metabolism; D023061:Gene Order; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D009876:Operon; D011485:Protein Binding; D016415:Sequence Alignment; D014169:Transformation, Bacterial
chemical_list: D004269:DNA, Bacterial; D003851:Deoxyribonucleases; D000251:Adenosine Triphosphatases