Topoisomerase Procaryotic

Addition of Mg to these complexes leads to their dissociation. However, addition of alkali leads to the appearance of cleaved DNA molecules having protein covalently attached to the DNA (see below). This enzyme also forms complexes with relaxed duplex DNA but these complexes are not dissociated by the addition of Mg and cleavage of the DNA is not found when alkali is added (Liu and Wang, 1979). Complexes between procaryotic topoisomerase I and DNA can also be detected by their retention on nitrocellulose filters. Using this method the preferential binding of the enzyme to an internal single-strand break in a linear duplex DNA has been shown (Dean et al., 1982 Dean and Cozzarelli, 1985). 

Although the DNA-binding properties of several procaryotic topoisomerases have been well characterized, little information is currently available concerning eucaryotic enzymes. Some eucaryotic topoisomerases may be intimately associated with other nuclear proteins HeLa topoisomerases I and II have been found to be associated with chromatin (Javaherian and Liu, 1983 Liu et al., 1983b). HeLa topoisomerase I has been shown to bind to the nonhistone protein HMG17, which also stimulates DNA catenation by the enzyme (Javaherian and Liu, 1983 Tse et al., 1984). It has been suggested that type II topoisomerase is an important component of the chromosomal scaffold in interphase nuclei and mitotic chromosomes from chicken cell lines (Earnshaw et al., 1985). In addition, an ATP-dependent topoisomerase has been found associated with several other enzymes of DNA metabolism in a complex (termed the replitase complex) isolated from the nuclei of Chinese hamster embryo fibroblast cells (Noguchi et al., 1983). 

Nalidixic acid and all its analogs act by selectively inhibiting bacterial DNA synthesis. The actual biochemical target of the quinolones is the enzyme DNA-gyrase, a topoisomerase enzyme found only in procaryotic cells. This accounts for the selective toxicity of these drugs. 

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