Reverse gyrase binding to DNA alters the double helix structure and produces single-strand cleavage in the absence of ATP.

Jaxel, Christine; Nadal, Marc; Mirambeau, Gilles; Forterre, Patrick; Takahashi, Miho; Duguet, Michel

doi:10.1002/j.1460-2075.1989.tb08466.x

Cited by 54 publications

(53 citation statements)

References 21 publications

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“…Together with previous studies using plasmids (7,9,20), these experiments with oligonucleotides further support the idea that the temperature dependence of the positive supercoiling reaction reflects the need for reverse gyrase to bind to locally denatured regions of the substrate.…”

Section: Dna Binding Behavior-supporting

confidence: 79%

“…1), similar to the S. acidocaldarius and Desulfurococcus amylolyticus enzymes (10,18,19). Numerous studies indicate that this temperature dependence reflects the need for singlestranded regions in the plasmid substrate (7,9,20). At elevated temperature, the DNA strands of a duplex tend to separate locally, especially when the DNA is negatively supercoiled, and this allows reverse gyrase to gain access to the DNA.…”

Section: Overexpression and Properties Of Recombinant Reversementioning

confidence: 82%

“…A contrasting explanation for the unidirectionality is provided by the "domain model" (6). Reverse gyrase is known to unwind DNA locally upon binding (9,21), and the domain model stipulates that the enzyme can isolate this unwound DNA topologically from the rest of the substrate. In DNA that is topologically constrained as a whole, such as a plasmid, the creation of unwound DNA at the protein binding site creates compensatory overwinding elsewhere.…”

Section: Fig 8 Topoisomerization Activity Of Reverse Gyrase Mutantmentioning

confidence: 99%

“…This was the end point of the reaction, because extending the incubation beyond 30 min did not lead to further relaxation (data not shown). The reaction was cooled on ice, causing the dissociation of protein from the plasmid (9). Then an excess of inactive reverse gyrase mutant was added to the reaction, and the 80°C incubation resumed.…”

Section: Fig 8 Topoisomerization Activity Of Reverse Gyrase Mutantmentioning

confidence: 99%

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Studies of a Positive Supercoiling Machine

Rodríguez

2002

Journal of Biological Chemistry

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Reverse gyrase, the only topoisomerase known to positively supercoil DNA, has an N-terminal ATPase domain that drives the activity of a topoisomerase domain. This study shows that the N-terminal domain represses topoisomerase activity in the absence of nucleotide, and nucleotide binding is sufficient to relieve the repression. A "latch" region in the N-terminal part was observed to close over the topoisomerase domain in the reverse gyrase crystal structure. Mutants lacking all or part of the latch relax DNA in the absence of nucleotide, indicating that this region mediates topoisomerase repression. The mutants also show altered DNA-dependent ATPase activity, suggesting that the latch may be involved in coupling nucleotide hydrolysis to supercoiling. It is not required for this process, however, because the mutants can still positively supercoil DNA. Nucleotide hydrolysis is essential to the specificity of reverse gyrase for increasing the linking number of DNA. Although with ATP the enzyme performs strand passage always toward increasing linking number, it can increase or decrease the linking number in the presence of a nonhydrolyzable ATP analog. This suggests that the mechanism of reverse gyrase is best described by a combination of recently proposed models.Topoisomerases participate in practically every DNA transaction, including transcription, replication, recombination, and chromosomal segregation (1). They change the topological state of DNA by altering its linking number, i.e. the number of times that one strand of the double helix crosses the other. They use a three-step mechanism of cleavage, strand passage, and religation. In the first step, a nucleophilic tyrosine attacks the phosphodiester backbone, cleaving the DNA and leading to a covalent intermediate of protein-DNA termed the "cleavage complex." Type I topoisomerases cleave one strand of the duplex; type II enzymes cleave both strands. The enzyme, covalently attached to the cut DNA, separates the free ends of the cleaved strand(s) and allows the other strand of the duplex (type I), or another region of duplex (type II), to pass through this gap. The protein then reseals the backbone of the cleaved DNA and releases the product.Although all topoisomerases can relax supercoiled DNA, only prokaryotic gyrase is able to introduce negative supercoils in DNA. This type II topoisomerase uses the energy of ATP hydrolysis to reduce the linking number, leading to an underwound, or negatively supercoiled, product. Until recently, gyrase was the only topoisomerase known to supercoil DNA. Later, another supercoiling enzyme was discovered and named reverse gyrase, because it increases the linking number, leading to overwound, or positively supercoiled, product (2, 3). Reverse gyrase is the only topoisomerase known to overwind DNA. So far it has been found exclusively in hyperthermophiles, organisms that live above ϳ80°C (4, 5). The role of the protein in vivo remains unclear (6); one suggestion is that it rewinds the DNA strands in regions of the chromosome that...

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Section: Dna Binding Behavior-supporting

confidence: 79%

Section: Overexpression and Properties Of Recombinant Reversementioning

confidence: 82%

Section: Fig 8 Topoisomerization Activity Of Reverse Gyrase Mutantmentioning

confidence: 99%

Section: Fig 8 Topoisomerization Activity Of Reverse Gyrase Mutantmentioning

confidence: 99%

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Studies of a Positive Supercoiling Machine

Rodríguez

2002

Journal of Biological Chemistry

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“…The first reverse gyrase characterized was isolated from the hyperthermophilic archaeum Sulfolobus acidocaldarius (23,33). Mechanistic studies showed that it is transiently linked to the DNA by a 5Ј phosphotyrosyl bond (22,24), classifying it in the type I 5Ј topoisomerase family as proposed by Roca (38). Sequence analysis further showed that it is a single polypeptide containing putative helicase and topoisomerase domains located in the amino-and carboxy-terminal, respectively, parts of the protein (9).…”

mentioning

confidence: 99%

Reverse gyrase from hyperthermophiles: probable transfer of a thermoadaptation trait from Archaea to Bacteria

et al. 2000

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The hyperthermophilic bacterium Thermotoga maritima MSB8 possesses a reverse gyrase whose enzymatic properties are very similar to those of archaeal reverse gyrases. It catalyzes the positive supercoiling of the DNA in an Mg 2؉ -and ATP-dependent process. Its optimal temperature of activity is around 90°C, and it is highly thermostable. We have cloned and DNA sequenced the corresponding gene (T. maritima topR). This is the first report describing the analysis of a gene encoding a reverse gyrase in bacteria. The T. maritima topR gene codes for a protein of 1,104 amino acids with a deduced molecular weight of 128,259, a value in agreement with that estimated from the denaturing gel electrophoresis of the purified enzyme. Like its archaeal homologs, the T. maritima reverse gyrase exhibits helicase and topoisomerase domains, and its sequence matches very well the consensus sequence for six reverse gyrases now available. Phylogenetic analysis shows that all reverse gyrases, including the T. maritima enzyme, form a very homogeneous group, distinct from the type I 5 topoisomerases of the TopA subfamily, for which we have previously isolated a representative gene in T. What are the molecular mechanisms involved in the adaptation of life to elevated temperatures? In terms of DNA dynamics, part of the answer was provided by the discovery in thermophilic organisms of a particular topoisomerase, the reverse gyrase, that modifies the topological state of DNA by introducing positive supercoils in an ATP-dependent process (14). It was suggested that overlinking could compensate for the effect of temperature on DNA structure (16). The enzyme is widely distributed in thermophilic archaea (6,8). The first reverse gyrase characterized was isolated from the hyperthermophilic archaeum Sulfolobus acidocaldarius (23,33). Mechanistic studies showed that it is transiently linked to the DNA by a 5Ј phosphotyrosyl bond (22,24), classifying it in the type I 5Ј topoisomerase family as proposed by Roca (38). Sequence analysis further showed that it is a single polypeptide containing putative helicase and topoisomerase domains located in the amino-and carboxy-terminal, respectively, parts of the protein (9). The helicase domain exhibits motifs found in DNA and RNA helicases, and the topoisomerase domain exhibits a significant similarity with the 5Ј topoisomerase I (protein ) from Escherichia coli. From all of these data, a mechanism of reverse gyration involving the concerted action of two such domains was proposed (9, 14).To date, four other archaeal reverse gyrase genes have been sequenced: from Sulfolobus shibatae (21), Methanopyrus kandleri (26), Pyrococcus furiosus (3), and Methanococcus jannaschii (7). A comparative analysis of reverse gyrases from two members of the order Sulfolobales (S. acidocaldarius and S. shibatae) and M. kandleri with the other type I topoisomerases of the 5Ј family (21) showed that the reverse gyrases constitute a new group within this family distinct from the previously described TopA and TopB groups, represent...

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Introduction and Historical Perspective

Forterre

2011

Cancer Drug Discovery and Development

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Reverse gyrase binding to DNA alters the double helix structure and produces single-strand cleavage in the absence of ATP.

Cited by 54 publications

References 21 publications

Studies of a Positive Supercoiling Machine

Studies of a Positive Supercoiling Machine

Reverse gyrase from hyperthermophiles: probable transfer of a thermoadaptation trait from Archaea to Bacteria

Introduction and Historical Perspective

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