Footprinting of yeast DNA topoisomerase II lysyl side chains involved in substrate binding and interdomainal interactions

被引:27
作者
Li, W [1 ]
Wang, JC [1 ]
机构
[1] Harvard Univ, Dept Mol & Cellular Biol, Cambridge, MA 02138 USA
关键词
D O I
10.1074/jbc.272.49.31190
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Footprinting of yeast DNA topoisomerase II and its NH2- and COOH-terminal truncation derivatives was carried out to map the locations of lysyl side chains that are involved in enzyme-DNA interaction, in the binding of ATP, or in interaction between domains of the same enzyme molecule, Several conclusions were drawn based on these measurements and the crystal structures of a 92-kDa fragment of the yeast enzyme and a 43-kDa fragment of Escherichia coli gyrase B-subunit, First, the footprinting results support the model previously in ferred from the 92-kDa fragment crystal structure that the main site of DNA binding is comprised of a pair of semicircular grooves, Second, the binding of a nonhydrolyzable ATP analog to the yeast enzyme appears to affect citraconylation at a minimum of six lysines in the ATPase domain of each polypeptide, Two of these lysines are probably involved in contacting the nucleotide directly, and one probably becomes buried when the two ATPase domains of a dimeric enzyme come into contact upon ATP binding; for the others, changes in lysine reactivity appear to reflect allosteric changes following ATP binding, Third, from a comparison of the footprint of the intact enzyme and those of the truncated polypeptides comprised of either the NH2- or the COOH-terminal half of the intact polypeptide, it appears that there are few contacts between the NH2- and COOH-terminal half of yeast DNA topoisomerase II.
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页码:31190 / 31195
页数:6
相关论文
共 28 条
  • [1] NUCLEOTIDE-BINDING TO THE 43-KILODALTON N-TERMINAL FRAGMENT OF THE DNA GYRASE-B PROTEIN
    ALI, JA
    ORPHANIDES, G
    MAXWELL, A
    [J]. BIOCHEMISTRY, 1995, 34 (30) : 9801 - 9808
  • [2] Study of yeast DNA topoisomerase II and its truncation derivatives by transmission electron microscopy
    Benedetti, P
    Silvestri, A
    Fiorani, P
    Wang, JC
    [J]. JOURNAL OF BIOLOGICAL CHEMISTRY, 1997, 272 (18) : 12132 - 12137
  • [3] Structure and mechanism of DNA topoisomerase II
    Berger, JM
    Gamblin, SJ
    Harrison, SC
    Wang, JC
    [J]. NATURE, 1996, 379 (6562) : 225 - 232
  • [4] BERGER JM, 1995, THESIS HARVARD U
  • [5] Caron P R, 1994, Adv Pharmacol, V29B, P271
  • [6] DNA SUPERCOILING INVIVO
    GIAEVER, GN
    SNYDER, L
    WANG, JC
    [J]. BIOPHYSICAL CHEMISTRY, 1988, 29 (1-2) : 7 - 15
  • [7] GOTO T, 1984, CELL, V36, P1073, DOI 10.1016/0092-8674(84)90057-6
  • [8] GOTO T, 1982, J BIOL CHEM, V257, P5866
  • [9] Binding of the sigma(70) protein to the core subunits of Escherichia coli RNA polymerase, studied by iron-EDTA protein footprinting
    Greiner, DP
    Hughes, KA
    Gunasekera, AH
    Meares, CF
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1996, 93 (01) : 71 - 75
  • [10] Radiolytic protein surface mapping
    Greiner, DP
    Hughes, KA
    Meares, CF
    [J]. BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1996, 225 (03) : 1006 - 1008