ROLE OF HU PROTEINS IN FORMING AND CONSTRAINING SUPERCOILS OF CHROMOSOMAL DNA IN ESCHERICHIA-COLI

被引:44
作者
TANAKA, H [1 ]
YASUZAWA, K [1 ]
KOHNO, K [1 ]
GOSHIMA, N [1 ]
KANO, Y [1 ]
SAIKI, T [1 ]
IMAMOTO, F [1 ]
机构
[1] KYOTO PHARMACEUT UNIV,INST MOLEC & CELLULAR BIOL PHARMACEUT SCI,DEPT MOLEC GENET,YAMASHIMA KU,KYOTO 607,JAPAN
来源
MOLECULAR & GENERAL GENETICS | 1995年 / 248卷 / 05期
关键词
HU; HU-LIKE PROTEIN; DNA SUPERCOILING; DNA GYRASE;
D O I
10.1007/BF02423446
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
Induction of supercoiling in plasmid DNA by HU heterotypic and homotypic dimers, a mutant HU-2 (HupAN12), HBs and HB1 proteins with different DNA-binding affinities was investigated in vitro. The abilities of these proteins to induce supercoiling in DNA correlated with their affinities for DNA. Stoichiometrical analysis of HU heterodimers bound to DNA in the complex restraining the negative torsional tension of DNA showed that 12-13 dimers account for a single superhelical turn. The number of supercoils in the plasmid in vivo decreased on inhibition of DNA gyrase with coumermycin, reaching a steady-state level that indicated the existence of a compartment of restrained supercoils. The size of the restrained compartment was reduced in the absence of HU, indicating the participation of HU in constituting this fraction, and was larger on overproduction of HU-2 in the cells. An increased level of DNA gyrase, expressed from a plasmid carrying both gyr genes, in the cells did not compensate for the deficit of the restrained supercoils caused by HU deficiency, indicating seeming distinct and unrelated action of HU and DNA gyrase in introducing and constraining supercoiling of intracellular DNA.
引用
收藏
页码:518 / 526
页数:9
相关论文
共 39 条
  • [1] Bonnefoy E., Rouviere-Yaniv J., HU and IHF, two homologous histone-like proteins of Escherichia coli, from different protein DNA complexes with short DNA fragments, EMBO J, 10, pp. 687-696, (1991)
  • [2] Broyles S., Pettijohn D.E., Interaction of the E. coli HU protein with DNA: evidence for formation of nucleosome-like structures with altered DNA helical pitch, J Mol Biol, 187, pp. 47-60, (1986)
  • [3] Craig N., Nash H., E. coli integration factor binds to specific sites in DNA, Cell, 39, pp. 707-716, (1984)
  • [4] Drlica K., Control of bacterial DNA supercoiling, Mol Microbiol, 6, pp. 425-433, (1992)
  • [5] Drlica K., Rotiviere-Yaniv J., Histone-like proteins of bacteria, Microbiol Rev, 51, pp. 301-319, (1987)
  • [6] Flashner Y., Gralla J.D., DNA dynamic flexibility and protein recognition: differential stimulation by bacterial histone-like protein HU, Cell, 54, pp. 713-721, (1988)
  • [7] Freidman D.I., Integration host factor: a protein for all reasons, Cell, 55, pp. 545-554, (1988)
  • [8] Gamper H., Hearst J., A topological model for transcription based on unwinding angle analysis of E. coli. RNA polymerase binary, initiation and ternary complexes, Cell, 29, pp. 81-90, (1982)
  • [9] Goshima N., Kano Y., Imamoto F., Characterization of HU-like protein from Bifidobacterium longum, Biochimie, 72, pp. 207-212, (1990)
  • [10] Goshima N., Kohno K., Imamoto F., Kano Y., HU-1 mutants of Escherichia coli deficient in DNA binding, Gene, 96, pp. 141-145, (1990)