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Separate and Combined Biochemical Activities of the Subunits of a Naturally Split Reverse Gyrase

Overview
Journal J Biol Chem
Specialty Biochemistry
Date 2010 Oct 9
PMID 20929866
Citations 13
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Abstract

Reverse gyrase reanneals denatured DNA and induces positive supercoils in DNA, an activity that is critical for life at very high temperatures. Positive supercoiling occurs by a poorly understood mechanism involving the coordination of a topoisomerase domain and a helicase-like domain. In the parasitic archaeon Nanoarchaeum equitans, these domains occur as separate subunits. We express the subunits, and characterize them both in isolation and as a heterodimer. Each subunit tightly associates and interacts with the other. The topoisomerase subunit enhances the catalytic specificity of the DNA-dependent ATPase activity of the helicase-like subunit, and the helicase-like subunit inhibits the relaxation activity of the topoisomerase subunit while promoting positive supercoiling. DNA binding preference for both single- and double-stranded DNA is partitioned between the subunits. Based on a sensitive topological shift assay, the binding preference of helicase-like subunit for underwound DNA is modulated by its binding with ATP cofactor. These results provide new insight into the mechanism of positive supercoil induction by reverse gyrase.

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References
1.
Schoeffler A, Berger J . DNA topoisomerases: harnessing and constraining energy to govern chromosome topology. Q Rev Biophys. 2008; 41(1):41-101. DOI: 10.1017/S003358350800468X. View

2.
Rodriguez A . Studies of a positive supercoiling machine. Nucleotide hydrolysis and a multifunctional "latch" in the mechanism of reverse gyrase. J Biol Chem. 2002; 277(33):29865-73. DOI: 10.1074/jbc.M202853200. View

3.
Capp C, Wu J, Hsieh T . Drosophila RecQ4 has a 3'-5' DNA helicase activity that is essential for viability. J Biol Chem. 2009; 284(45):30845-52. PMC: 2781483. DOI: 10.1074/jbc.M109.008052. View

4.
Atomi H, Matsumi R, Imanaka T . Reverse gyrase is not a prerequisite for hyperthermophilic life. J Bacteriol. 2004; 186(14):4829-33. PMC: 438624. DOI: 10.1128/JB.186.14.4829-4833.2004. View

5.
Krah R, Kozyavkin S, Slesarev A, Gellert M . A two-subunit type I DNA topoisomerase (reverse gyrase) from an extreme hyperthermophile. Proc Natl Acad Sci U S A. 1996; 93(1):106-10. PMC: 40187. DOI: 10.1073/pnas.93.1.106. View