Characterization of the mycobacterial AdnAB DNA motor provides insights into the evolution of bacterial motor-nuclease machines.

The Journal of Biological Chemistry
Mihaela-Carmen Unciuleac, Stewart Shuman

Abstract

Mycobacterial AdnAB exemplifies a family of heterodimeric motor-nucleases involved in processing DNA double strand breaks (DSBs). The AdnA and AdnB subunits are each composed of an N-terminal UvrD-like motor domain and a C-terminal RecB-like nuclease module. Here we conducted a biochemical characterization of the AdnAB motor, using a nuclease-inactivated heterodimer. AdnAB is a vigorous single strand DNA (ssDNA)-dependent ATPase (k(cat) 415 s(-1)), and the affinity of the motor for the ssDNA cofactor increases 140-fold as DNA length is extended from 12 to 44 nucleotides. Using a streptavidin displacement assay, we demonstrate that AdnAB is a 3' --> 5' translocase on ssDNA. AdnAB binds stably to DSB ends. In the presence of ATP, the motor unwinds the DNA duplex without requiring an ssDNA loading strand. We integrate these findings into a model of DSB unwinding in which the "leading" AdnB and "lagging" AdnA motor domains track in tandem, 3' to 5', along the same DNA single strand. This contrasts with RecBCD, in which the RecB and RecD motors track in parallel along the two separated DNA single strands. The effects of 5' and 3' terminal obstacles on ssDNA cleavage by wild-type AdnAB suggest that the AdnA nuclease receives and proc...Continue Reading

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Citations

Sep 22, 2010·Quarterly Reviews of Biophysics·Wei Yang
Sep 21, 2010·Nature Structural & Molecular Biology·Colin G WuTimothy M Lohman
Jun 27, 2013·Proceedings of the National Academy of Sciences of the United States of America·Carolina CarrascoFernando Moreno-Herrero
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Nov 20, 2019·Proceedings of the National Academy of Sciences of the United States of America·Ning JiaStewart Shuman
Dec 4, 2012·Nature Reviews. Microbiology·Dale B Wigley
Apr 11, 2021·Proceedings of the National Academy of Sciences of the United States of America·Mihaela-Carmen UnciuleacStewart Shuman

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