Biopolym. Cell. 1987; 3(2):77-82.
Structure and Function of Biopolymers
Nuclear DNA topoisomerase from the Zea mays embryos
1Karpenchuk K. G., 1Rudenko G. N.
  1. N. G. Kholodny Institute of Botany, Academy of Sciences of the Ukrainian SSR
    Kiev, USSR

Abstract

ATP-independent presumably type I DNA topoisomerase was partially purified from the nuclear extract of Zea mays mature embryos by separation in the polyethylene glycol-ammonium sulphate system with 2 M NaCl and by salt gradient elution from CM-Sepha-dex. The ability of enzyme to relax negative supercoils of plasmid pBR322 DNA was detected in the presence of EDTA, being however markedly stimulated by the magnesium ions and polyamines (spermidine and cadaverine). The reaction proceeded via either processive or distributive mechanism depending on the ionic strength of the enzyme assay mixture. Novobiocin and nalidixic acid (both at 100 u.g/ml) did not inhibit the enzyme activity. Modification with p-hydroxymercuribenzoate or iodoacetamide completely abolished the DNA topoisomerase activity, thus indicating the presence of essential cysteine residues in the enzyme molecules. The DNA topoisomerase activity was found to be completely depressed by the preparation of potent inhibitor of polynucleotide-binding proteins – aurintricarboxylic acid in partially polymerized form. Magnesium ions partially restored the DNA topoisomerase activity in the presence of aurintricarboxylic acid.

References

[1] Gellert M. DNA topoisomerases. Annu Rev Biochem. 1981;50:879-910.
[2] Kafiani KA, Bronshtein IB, Gromova II, Shakhbazian GK, Timofeev AV. DNA-topoisomerases of animal cells. Mol Biol (Mosk). 1985;19(2):438-49.
[3] Bauer WR. Structure and reactions of closed duplex DNA. Annu Rev Biophys Bioeng. 1978;7:287-313.
[4] Wang JC, Peck LJ, Becherer K. DNA supercoiling and its effects on DNA structure and function. Cold Spring Harb Symp Quant Biol. 1983;47 Pt 1:85-91.
[5] Wang IC. Unwinding of the promoter and the modulation of transcription by DNA supercoiling. Promoters. Structure and Function. Eds R. L. Rodriguez, M. J. Chamberlin - New York: Praeger Publ., 1982:229-241.
[6] Stirdivant SM, Crossland LD, Bogorad L. DNA supercoiling affects in vitro transcription of two maize chloroplast genes differently. Proc Natl Acad Sci U S A. 1985;82(15):4886-90.
[7] Richardson SM, Higgins CF, Lilley DM. The genetic control of DNA supercoiling in Salmonella typhimurium. EMBO J. 1984;3(8):1745-52.
[8] Mirkin SM, Zaitsev EN, Panyutin IG, Lyamichev VI. Native supercoiling of DNA: the effects of DNA gyrase and omega protein in E. coli. Mol Gen Genet. 1984;196(3):508-12.
[9] Goto T, Laipis P, Wang JC. The purification and characterization of DNA topoisomerases I and II of the yeast Saccharomyces cerevisiae. J Biol Chem. 1984;259(16):10422-9.
[10] Dynan WS, Jendrisak JJ, Hager DA, Burgess RR. Purification and characterization of wheat germ DNA topoisomerase I (nicking-closing enzyme). J Biol Chem. 1981;256(11):5860-5.
[11] Siedlecki J, Zimmermann W, Weissbach A. Characterization of a prokaryotic topoisomerase I activity in chloroplast extracts from spinach. Nucleic Acids Res. 1983;11(5):1523-36.
[12] Fukata H, Fukasawa H. Isolation and partial characterization of two distinct DNA topoisomerases from cauliflower inflorescence. J Biochem. 1982;91(4):1337-42.
[13] Reeves R. Transcriptionally active chromatin. Biochim Biophys Acta. 1984;782(4):343-93.
[14] Fleischmann G, Pflugfelder G, Steiner EK, Javaherian K, Howard GC, Wang JC, Elgin SC. Drosophila DNA topoisomerase I is associated with transcriptionally active regions of the genome. Proc Natl Acad Sci U S A. 1984;81(22):6958-62.
[15] McConaughy BL, Young LS, Champoux JJ. The effect of salt on the binding of the eucaryotic DNA nicking-closing enzyme to DNA and chromatin. Biochim Biophys Acta. 1981;655(1):1-8.
[16] Blumenthal T, Landers TA. The inhibition of nucleic acid-binding proteins by aurintricarboxylic acid. Biochem Biophys Res Commun. 1973;55(3):680-8.
[17] Gonzalez RG, Blackburn BJ, Schleich T. Fractionation and structural elucidation of the active components of aurintricarboxylic acid, a potent inhibitor of protein nucleic acid interactions. Biochim Biophys Acta. 1979;562(3):534-45.
[18] Gonzalez RG, Haxo RS, Schleich T. Mechanism of action of polymeric aurintricarboxylic acid, a potent inhibitor of protein--nucleic acid interactions. Biochemistry. 1980;19(18):4299-303.
[19] Shen LL, Pernet AG. Mechanism of inhibition of DNA gyrase by analogues of nalidixic acid: the target of the drugs is DNA. Proc Natl Acad Sci U S A. 1985;82(2):307-11.
[20] Tabor CW, Tabor H. 1,4-Diaminobutane (putrescine), spermidine, and spermine. Annu Rev Biochem. 1976;45:285-306.
[21] Klevan , Tse Y-C. Chemical modification of essential tyrosine residues in DNA topoisomerases. Biochim Biophys Acta. 1983; 745(2):175-80.
[22] Ishii K, Katase A, Andoh T, Seno N. Inhibition of topoisomerase I by heparin. Biochem Biophys Res Commun. 1982;104(2):541-7.
[23] Liu LF, Miller KG. Eukaryotic DNA topoisomerases: two forms of type I DNA topoisomerases from HeLa cell nuclei. Proc Natl Acad Sci U S A. 1981;78(6):3487-91.