Literature summary for 5.6.2.2 extracted from

Buhler, C.; Gadelle, D.; Forterre, P.; Wang, J.C.; Bergerat, A.
Reconstitution of DNA topoisomerase VI of the thermophilic archaeon Sulfolobus shibatae from subunits separately overexpressed in Escherichia coli (1998), Nucleic Acids Res., 26, 5157-5162.

Cloned(Commentary)

Cloned (Comment)	Organism
overexpression in Escherichia coli. The A subunit exhibits clusters of arginines encoded by rare codons in Escherichia coli. The expression of this protein requires the coexpression of the minor Escherichia coli arginyl tRNA which reads AGG and AGA codons. The A subunit expressed in Escherichia coli is obtained from inclusion bodies after denaturation and renaturation. The B subunit is overexpressed in Escherichia coli and purified in soluble form. When purified B subunit is added to the renatured A subunit, ATP-dependent relaxation and decatenation activities of the hyperthermophilic DNA topoisomerase are reconstituted. The reconstituted recombinant enzyme exhibits a specific activity similar to the enzyme purified from Sulfolobus shibatae. It catalyzes transient double-strain cleavage of DNA and becomes covalently attached to the ends of the cleaved DNA	Saccharolobus shibatae

Cloned (Comment)

Organism

overexpression in Escherichia coli. The A subunit exhibits clusters of arginines encoded by rare codons in Escherichia coli. The expression of this protein requires the coexpression of the minor Escherichia coli arginyl tRNA which reads AGG and AGA codons. The A subunit expressed in Escherichia coli is obtained from inclusion bodies after denaturation and renaturation. The B subunit is overexpressed in Escherichia coli and purified in soluble form. When purified B subunit is added to the renatured A subunit, ATP-dependent relaxation and decatenation activities of the hyperthermophilic DNA topoisomerase are reconstituted. The reconstituted recombinant enzyme exhibits a specific activity similar to the enzyme purified from Sulfolobus shibatae. It catalyzes transient double-strain cleavage of DNA and becomes covalently attached to the ends of the cleaved DNA

Saccharolobus shibatae

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
47000	-	2 * 47000 + 2 * 60000	Saccharolobus shibatae
60000	-	2 * 47000 + 2 * 60000	Saccharolobus shibatae

Organism

Organism	UniProt	Comment	Textmining
Saccharolobus shibatae	O05208 and O05207	type II DNA topoisomerase VI subunit A and B	-

Purification (Commentary)

Purification (Comment)	Organism
purification of the two recombinant subunits of Sulfolobus shibatae Topo IV from Escherichia coli	Saccharolobus shibatae

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
DNA + ATP	DNA topoisomerase VI relaxes both negatively and positively supercoiled DNA in the presence of ATP and has no DNA supercoiling activity	Saccharolobus shibatae	?	-	?

Subunits

Subunits	Comment	Organism
tetramer	2 * 47000 + 2 * 60000	Saccharolobus shibatae

Synonyms

Synonyms	Comment	Organism
DNA topoisomerase VI	-	Saccharolobus shibatae