{"doi":"10.1111/j.1365-2958.2006.05068.x","title":"The intrinsic ATPase activity of <i>Mycobacterium tuberculosis</i> DnaA promotes rapid oligomerization of DnaA on <i>oriC</i>","abstract":"<jats:title>Summary</jats:title><jats:p>Oligomerization of the initiator protein, DnaA, on the origin of replication (<jats:italic>oriC</jats:italic>) is crucial for initiation of DNA replication<jats:italic>.</jats:italic> Studies in <jats:italic>Escherichia coli</jats:italic> (Gram‐negative) have revealed that binding of DnaA to ATP, but not hydrolysis of ATP, is sufficient to promote DnaA binding, oligomerization and DNA strand separation. To begin understanding the initial events involved in the initiation of DNA replication in <jats:italic>Mycobacterium tuberculosis</jats:italic> (Gram‐positive), we investigated interactions of <jats:italic>M. tuberculosis</jats:italic> DnaA (DnaA<jats:sub>TB</jats:sub>) with <jats:italic>oriC</jats:italic> using surface plasmon resonance in the presence of ATP and ADP. We provide evidence that, in contrast to what is observed in <jats:italic>E. coli</jats:italic>, ATPase activity of DnaA<jats:sub>TB</jats:sub> promoted rapid oligomerization on <jats:italic>oriC.</jats:italic> In support, we found that a recombinant mutant DnaA<jats:sub>TB</jats:sub> proficient in binding to ATP, but deficient in ATPase activity, did not oligomerize as rapidly. The corresponding mutation in the <jats:italic>dnaA</jats:italic> gene of <jats:italic>M. tuberculosis</jats:italic> resulted in non‐viability, presumably due to a defect in <jats:italic>oriC</jats:italic>–DnaA interactions. Dimethy sulphate (DMS) footprinting experiments revealed that DnaA<jats:sub>TB</jats:sub> bound to DnaA boxes similarly with ATP or ADP. DnaA<jats:sub>TB</jats:sub> binding to individual DnaA boxes revealed that rapid oligomerization on <jats:italic>oriC</jats:italic> is triggered only after the initial interaction of DnaA with individual DnaA boxes. We propose that ATPase activity enables the DnaA protomers on <jats:italic>oriC</jats:italic> to rapidly form oligomeric complexes competent for replication initiation.</jats:p>","journal":"Molecular Microbiology","year":2006,"id":675160,"datarank":0.5533319181170905,"base_score":3.6888794541139363,"endowment":3.6888794541139363,"self_citation_contribution":0.5533319181170905,"citation_network_contribution":0.0,"self_endowment_contribution":0.5533319181170905,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":39,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":null,"is_data_producer":false,"deposit_databanks":null,"is_oa":false,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":null,"fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":1764090,"name":"Meredith Moomey","orcid":null,"position":1,"is_corresponding":false},{"id":1466185,"name":"Pierre F. Neuenschwander","orcid":"0009-0003-6739-7516","position":2,"is_corresponding":false},{"id":1764091,"name":"Syed Muniruzzaman","orcid":null,"position":3,"is_corresponding":false},{"id":1764092,"name":"Kohji Yamamoto","orcid":null,"position":4,"is_corresponding":false},{"id":1764093,"name":"Julia E. Grimwade","orcid":null,"position":5,"is_corresponding":false},{"id":1764094,"name":"Malini Rajagopalan","orcid":null,"position":6,"is_corresponding":false},{"id":1764089,"name":"Murty V. V. S. 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To begin understanding the initial events involved in the initiation of DNA replication in <jats:italic>Mycobacterium tuberculosis</jats:italic> (Gram‐positive), we investigated interactions of <jats:italic>M. tuberculosis</jats:italic> DnaA (DnaA<jats:sub>TB</jats:sub>) with <jats:italic>oriC</jats:italic> using surface plasmon resonance in the presence of ATP and ADP. We provide evidence that, in contrast to what is observed in <jats:italic>E. coli</jats:italic>, ATPase activity of DnaA<jats:sub>TB</jats:sub> promoted rapid oligomerization on <jats:italic>oriC.</jats:italic> In support, we found that a recombinant mutant DnaA<jats:sub>TB</jats:sub> proficient in binding to ATP, but deficient in ATPase activity, did not oligomerize as rapidly. The corresponding mutation in the <jats:italic>dnaA</jats:italic> gene of <jats:italic>M. tuberculosis</jats:italic> resulted in non‐viability, presumably due to a defect in <jats:italic>oriC</jats:italic>–DnaA interactions. Dimethy sulphate (DMS) footprinting experiments revealed that DnaA<jats:sub>TB</jats:sub> bound to DnaA boxes similarly with ATP or ADP. DnaA<jats:sub>TB</jats:sub> binding to individual DnaA boxes revealed that rapid oligomerization on <jats:italic>oriC</jats:italic> is triggered only after the initial interaction of DnaA with individual DnaA boxes. 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