title: The essential transfer protein TraM binds to DNA as a tetramer.
abstract: The TraM proteins encoded by F-like plasmids are sequence specific DNA binding proteins that are essential for conjugative DNA transfer. We investigated the quarternary structure and the DNA binding properties of the TraM wild-type protein of the resistance plasmid R1 and two mutant forms thereof. Size-exclusion chromatography and differential scanning calorimetry showed that purified TraM protein (amino acids 2-127) forms stable tetramers in solution. A truncated version of the protein termed TraMM26 (amino acids 2-56) forms dimers. Thus, the dimerization and tetramerization domains can be assigned to the N-terminal and C-terminal domains of TraM, respectively. Further analyses using chemical cross-linking and light scattering corroborated the preferentially tetrameric nature of the protein but also suggest that TraM has a tendency to form higher aggregates. Band-shift and fluorescence spectroscopy investigations of TraM-DNA complexes revealed that the TraM protein is also tetrameric when bound to its minimal DNA binding site. The deduced binding constant in the range of 10(8) M(-1) demonstrated a very strong binding of TraM to its preferred DNA sequence. Secondary structure analysis based on CD measurements showed that TraM is mainly alpha-helical with a significant increase in alpha-helicity (48 to 58%) upon DNA-binding, indicating an induced fit mechanism.
authors.affiliation: Institut für Physikalische Chemie, Karl-Franzens-Universität Graz, Heinrichstr. 28, A-8010 Graz, Austria.
authors.forename: P
authors.initials: P
authors.lastname: Verdino
authors.forename: W
authors.initials: W
authors.lastname: Keller
authors.forename: H
authors.initials: H
authors.lastname: Strohmaier
authors.forename: K
authors.initials: K
authors.lastname: Bischof
authors.forename: H
authors.initials: H
authors.lastname: Lindner
authors.forename: G
authors.initials: G
authors.lastname: Koraimann
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D002942:Circular Dichroism; D004247:DNA / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000737:chemistry*; D017510:Protein Folding; D017433:Protein Structure, Secondary
chemical_list: D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; C074281:TraM protein, bacterial; D004247:DNA