title: DNA binding specificity of the replication initiator protein, DnaA from Helicobacter pylori.
abstract: The key protein in the initiation of Helicobacter pylori chromosome replication, DnaA, has been characterized. The amount of the DnaA protein was estimated to be approximately 3000 molecules per single cell; a large part of the protein was found in the inner membrane. The H.pylori DnaA protein has been analysed using in vitro (gel retardation assay and surface plasmon resonance (SPR)) as well as in silico (comparative computer modeling) studies. DnaA binds a single DnaA box as a monomer, while binding to the fragment containing several DnaA box motifs, the oriC region, leads to the formation of high molecular mass nucleoprotein complexes. In comparison with the Escherichia coli DnaA, the H.pylori DnaA protein exhibits lower DNA-binding specificity; however, it prefers oriC over non-box DNA fragments. As determined by gel retardation techniques, the H.pylori DnaA binds with a moderate level of affinity to its origin of replication (4nM). Comparative computer modelling showed that there are nine residues within the binding domain which are possible determinants of the reduced H.pylori DnaA specificity. Of these, the most interesting is probably the triad PTL; all three residues show significant divergence from the consensus, and Thr398 is the most divergent residue of all.
authors.affiliation: Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Wrocław, Poland.
authors.forename: Anna
authors.initials: A
authors.lastname: Zawilak
authors.forename: Marcus C
authors.initials: MC
authors.lastname: Durrant
authors.forename: Piotr
authors.initials: P
authors.lastname: Jakimowicz
authors.forename: Steffen
authors.initials: S
authors.lastname: Backert
authors.forename: Jolanta
authors.initials: J
authors.lastname: Zakrzewska-Czerwińska
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism*; D001483:Base Sequence; D017931:DNA Primers; D004269:DNA, Bacterial / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000378:metabolism*; D016480:Helicobacter pylori / Q000378:metabolism*; D008958:Models, Molecular; D011485:Protein Binding; D020349:Surface Plasmon Resonance
chemical_list: D001426:Bacterial Proteins; D017931:DNA Primers; D004269:DNA, Bacterial; D004268:DNA-Binding Proteins; C066312:DnaA protein, Bacteria