The latch modulates nucleotide and DNA binding to the helicase-like domain of Thermotoga maritima reverse gyrase and is required for positive DNA supercoiling
Open Access
- 4 November 2010
- journal article
- research article
- Published by Oxford University Press (OUP) in Nucleic Acids Research
- Vol. 39 (5), 1789-1800
- https://doi.org/10.1093/nar/gkq1048
Abstract
Reverse gyrase is the only topoisomerase that can introduce positive supercoils into DNA in an ATP-dependent process. It has a modular structure and harnesses a helicase-like domain to support a topoisomerase activity, thereby creating the unique function of positive DNA supercoiling. The isolated topoisomerase domain can relax negatively supercoiled DNA, an activity that is suppressed in reverse gyrase. The isolated helicase-like domain is a nucleotide-dependent switch that is attenuated by the topoisomerase domain. Inter-domain communication thus appears central for the functional cooperation of the two domains. The latch, an insertion into the helicase-like domain, has been suggested as an important element in coordinating their activities. Here, we have dissected the influence of the latch on nucleotide and DNA binding to the helicase-like domain, and on DNA supercoiling by reverse gyrase. We find that the latch is required for positive DNA supercoiling. It is crucial for the cooperativity of DNA and nucleotide binding to the helicase-like domain. The latch contributes to DNA binding, and affects the preference of reverse gyrase for ssDNA. Thus, the latch coordinates the individual domain activities by modulating the helicase-like domain, and by communicating changes in the nucleotide state to the topoisomerase domain.Keywords
This publication has 24 references indexed in Scilit:
- Features and development of CootActa Crystallographica Section D-Biological Crystallography, 2010
- The reverse gyrase helicase-like domain is a nucleotide-dependent switch that is attenuated by the topoisomerase domainNucleic Acids Research, 2008
- Adenosine 5′-O-(3-thio)triphosphate (ATPγS) Promotes Positive Supercoiling of DNA by T. maritima Reverse GyraseJournal of Molecular Biology, 2007
- Reverse Gyrase Functions as a DNA RenaturaseJournal of Biological Chemistry, 2006
- Investigating the Role of the Latch in the Positive Supercoiling Mechanism of Reverse GyraseBiochemistry, 2003
- Studies of a Positive Supercoiling MachineJournal of Biological Chemistry, 2002
- Reverse Gyrase, the Two Domains Intimately Cooperate to Promote Positive SupercoilingJournal of Biological Chemistry, 2000
- The CCP4 suite: programs for protein crystallographyActa Crystallographica Section D-Biological Crystallography, 1994
- Reverse gyrase—a topoisomerase which introduces positive superhelical turns into DNANature, 1984
- New ribose-modified fluorescent analogs of adenine and guanine nucleotides available as subtrates for various enzymesBiochimica et Biophysica Acta (BBA) - Protein Structure and Molecular Enzymology, 1983