Identification of the primary metal ion-activation sites of the diphtheria tox repressor by X-ray crystallography and site-directed mutational analysis

Nature Structural Biology

Volumn 3, Issue 4, 1996, Pages 382-387

  Ding, X ^a   Zeng, H ^b   Schiering, N ^b,c   Ringe, D ^a   Murphy, J R ^b  

^a BRANDEIS UNIVERSITY   (United States)

^b BOSTON MEDICAL CENTER   (United States)

^c PFIZER INC   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

DIPHTHERIA TOXIN; METAL ION; NICKEL; REPRESSOR PROTEIN;

ARTICLE; BACTERIAL GENE; CORYNEBACTERIUM DIPHTHERIAE; GENE EXPRESSION; PRIORITY JOURNAL; PROTEIN DNA BINDING; SITE DIRECTED MUTAGENESIS; STRUCTURAL GENE; TOXIN STRUCTURE; X RAY CRYSTALLOGRAPHY;

BACTERIAL PROTEINS; BINDING SITES; CATIONS, DIVALENT; CORYNEBACTERIUM DIPHTHERIAE; CRYSTALLOGRAPHY, X-RAY; DNA MUTATIONAL ANALYSIS; DNA, BACTERIAL; DNA-BINDING PROTEINS; MODELS, MOLECULAR; MUTAGENESIS, SITE-DIRECTED; NICKEL; PROTEIN CONFORMATION;

BACTERIA (MICROORGANISMS); CORYNEBACTERIUM DIPHTHERIAE;

EID: 0029964556     PISSN: 10728368     EISSN: None     Source Type: Journal    
DOI: 10.1038/nsb0496-382     Document Type: Article

References (21)

1. Tao, X., Schiering, N., Zeng, H., Ringe, D. & Murphy, J.R. Iron, DtxR, and the regulation of diphtheria toxin expression. Molec. Microbiol. 14, 191-197 (1994).
2. Pappenheimer, A.M., Jr. The pathogenesis of diphtheria. Symp. Soc. Gen. Microbiol., 40-56 (1955).
3. Uchida, T., Gill, D.M. & Pappenheimer, A.M., Jr. Mutation in the structural gene for diphtheria toxin carried by temperate phage β. Nature 233, 8-11 (1971).
4. Buck, G.A., Gross, R.E., Wong, T.P., Lorea, T. & Groman, N.B. DNA relationships among some tox-bearing corynebacteriophages. Infect. Immun. 49, 679-684 (1985).
5. Boyd, J., Oza, M. & Murphy, J.R. Molecular cloning and DNA sequence analysis of an iron dependent diphtheria tax regluatory element (dtxK) from Corynebacterium diphtheriae. Proc. Natl. Acad. Sci. USA 87, 5968-5972 (1990).
6. Schmitt, M.P. & Holmes, R.K. Iron-dependent regulation of diphtheria toxin and siderophores expression by the cloned Corynebacterium diphtheriae repressor gene dtxK in C. diphtheriae. Infect. Immun. 59, 1899-1904 (1991).
7. Tao, X., Boyd, J. & Murphy, J.R. Specific binding of the diphtheria tox regulatory element DtxR to the tox operator requires divalent cations and a 9-base-pair interrupted palindromic sequence. Proc. Natl. Acad. Sci. USA 89, 5897-5901 (1992).
8. Schmitt, M.P. & Holmes, R.K. Analysis of diphtheria toxin repressor-operator interactions and characterization of a mutant repressor with decreased binding activity for divalent matais. Mol. Microbiol. 9, 173-181 (1993).
9. Tao, X., & Murphy, J.R. Determination of the DtxR consensus binding site by in vitro affinity selection. Proc. Natl. Acad. Sci. USA 91, 9646-9650 (1994).
10. Qiu, X. et al. Three-dimensional structure of the diphtheria toxin repressor in complex with divalent cation corepressors. Structure 3, 87-100 (1995).
11. Schiering, N. et al. Structures of the apo- and the metal ion activated forms of the diphtheria tox repressor from Corynebacterium diphtherias. Proc. Natl. Acad. Sci. USA 92, 9843-9850 (1995).
12. Wang, Z., Schmitt, M.P. & Holmes, R.K. Characterization of mutations that inactivate the diphtheria toxin repressor (dtxR). Infect. Immun. 62, 1600-1608 (1994).
13. Tao, X., Zeng, H. & Murphy, J.R. Transition metal ion activation of DNA binding by the diphtheria tox repressor requires the formation of stable homodimers. Proc. Natl. Acad. Sci. USA 92, 6803-6807 (1995).
14. Tao, X. & Murphy, J.R. Cysteine-102 is positioned in the metal ion activation site of the Corynebacterium diphtherias regulatory element DtxR. Proc. Natl. Acad. Sci. USA 90, 8524-8528 (1993).
15. Sanger, F., Nicklen, S. & Coulsen A.R. DNA sequencing with with chain terminating inhibitors. Proc. Natl. Acad. Sci. USA 74, 5463-5467 (1977).
16. Kraft, R., Tardiff, J., Krauter, K.S. & Leinwand, L.A. Using mini-prep plasmid DNA for sequencing double stranded template with sequenase. BioTechniques 6, 544-547 (1988).
17. Miller, J.H. Experiments in Molecular Genetics (Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, 1972)
18. Putnam, S.L. & Koch, A.L. Complications in the simplest cellular assay: Lysis of Escherichia coli for the assay of β-galactosidase. Anal. Biochem. 63, 350-360 (1975).
19. Brünger, A.T., Karplus, M. & Kuriyan, J. Crystallographic R-factor refinement by molecular dynamics. Science 235, 458-460 (1987).
20. Brünger, A.T. Crystallographic refinement by simulate annealing: application to a 2.8Å resolution structure of aspartate aminotransferase. J. Mol. Biol. 203, 803-816 (1988).
21. Jones, T.A., Zou, J.Y., Cowan, S.W. & Kjeldgaard. Improved methods for building protein models in electron density maps and the location of errors in these models. Acta Crystallogr. A47, 110-119 (1991).