Interaction Between Replication Initiator DnaA And DnbM In Mycobacterium Tuberculosis

Mycobacterium tuberculosis is one of the most important human pathogens,and very dangerous to human life and health.DNA replication initiation,one of the essential biological processes of bacteria,is also key to M.Tuberculosis.However,the regulation mechanisms on DNA replication remain unknown in M Tuberculosis.DnaA is a significant replication initiation protein,which plays critical role in replication initiation stage.Therefore,studies on the DnaA and its regulation will facilitate us to understand the unique DNA replication mechanisms of M.Tuberculosis and result in new potential drug targets.Library of M.Tuberculosis was screened with DnaA,and DnbM,a potential nucleoid-associated protein identified by our lab,was found to interact with DnaA.This interaction was further studied and the results are as follows:(1)The physcial interaction between DnaA and DnbM protein was confirmed by two hybrid assay,which is through DnaA N-terminal domain and DnbM C-ternimal domain.(2)SPR and pull-down assays further confirmed the physcial interaction between DnaA and DnbM protein.(3)Furthermore,the interaction between DnaA and DnbM was proved in vivo by Co-IP assay.(4)At last,we demonstrated that DnbM protein can supress DnaA binding oriC via the interaction benween DnaA and DnbM.In summary,we confirmed the interaction between DnaA and DnbM,and the interaction between initation factor DnaA and oriC with DnbM.The results will contribute to understand the mechanism of the DNA replication in M.tuberculosis,and can provide new insight for the treatment of tuberculosis.