DNA binding activity ofHelicobacter pyloriDnaB helicase: the role of the N-terminal domain in modulating DNA binding activities


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Abstract

Replicative helicases are major motor proteins essential for chromosomal DNA replication in prokaryotes. Usually hexameric in solution, their DNA binding property must have different roles at stages ranging from the loading onto a branched structure at initiation from the origin to the highly processive translocation during elongation. Here, we have analysed the DNA binding activity of Helicobacter pylori (Hp) replicative helicase, DnaB. The results indicate that while the C-terminal region is important for its DNA binding activity, the N-terminus appears to dampen the protein’s affinity for DNA. The masking activity of the N-terminus does not require ATP or hexamerization of HpDnaB and can be overcome by deleting the N-terminus. It can also be neutralized by engaging the N-terminus in an interaction with a partner like the C-terminus of DnaG primase. The inhibitory effect of the N-terminus on DNA binding activity is consistent with the 3D homology model of HpDnaB. Electron microscopy of the HpDnaB–ssDNA complex showed that HpDnaB preferentially bound at the ends of linear ssDNA and translocated along the DNA in the presence of ATP. These results provide an insight into the stimulatory and inhibitory effects of different regions of HpDnaB on DNA binding activities that may be central to the loading and translocation functions of DnaB helicases.Structured digital abstractdnaB and dnaB bind by molecular sieving (View interaction)dnaB and dnaB bind by circular dichroism (View interaction)In this study, we have analysed the DNA binding activity of H. pylori replicative helicase, DnaB. While the C-terminal region of HpDnaB is important for its DNA binding activity, the N-terminus masks the DNA binding activity profoundly. The masking effect can be overcome either by deleting the N-terminus or by interaction with a partner like the C-terminus of DnaG primase.

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