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PhD (laser physics)
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A whole genome approach to in vivo DNA-protein interactions in E. coli

Ming X. Wang* & George M. Church†

* Laboratory of Oncology Research, Research Division, Wills Eye Hospital and Jefferson Medical College of Thomas Jefferson University, Philadelphia, Pennsylvania 19107, USA
† Department of Genetics, Harvard Medical School and Howard Hughes Medical Institute, Boston, Massachusetts 02115, USA

THE increasingly rapid pace at which genomic DNA sequences are being determined has created a need for more efficient techniques to determine which parts of these sequences are bound in vivo by the proteins controlling processes such as gene expression, DNA replication and chromosomal mechanics. Here we describe a whole-genome approach to identify and characterize such DNA sequences. The method uses endogenous or artificially introduced methylases to methylate all genomic targets except those protected in vivo by protein or non-protein factors interfering with methylase action. These protected targets remain unmethylated in purified genomic DNA and are identified using methylation-sensitive restriction endonucleases. When the method was applied to the Escherichia coli genome, 0.1% of the endogenous adenine methyltransferase (Dam methylase) targets were found to be unmethylated. Five foreign methylases were examined by transfection. Database-matched DNA sequences flanking the in vivo -protected Dam sites all fell in the non-coding regions of seven E. coli operons (mtl, cdd, flh, gut, car, psp and fep). In the first four operons these DNA sequences closely matched the consensus sequence that binds to the cyclic AMP-receptor protein. The in vivo protection at the Dam site upstream of the car operon was correlated with a downregulation of car expression, as expected of a feedback represser-binding model.

References

1. Razin, A. et al. Nucleic Acids Res. 8, 1783−1792 (1980). | PubMed | ChemPort |
2. Geier, G. & Modrich, P. J. biol. Chem. 254, 1408−1413 (1979). | PubMed | ISI | ChemPort |
3. Blyn, L. B., Braaten, B. A. & Low, D. A. EMBO J. 9, 4045−4054 (1990). | PubMed | ISI | ChemPort |
4. Ringquist, S. & Smith, C. L. Proc. natn. Acad. Sci. U.S.A. 89, 4539−4543 (1992). | ChemPort |
5. Braaten, B. A. et al. Proc. natn. Acad. Sci. U.S.A. 89, 4250−4254 (1992). | ChemPort |
6. Campbell, J. L. & Kleckner, N. Gene 74, 189−190 (1988). | Article | PubMed | ISI | ChemPort |
7. Jaworski, A. et al. Science 238, 773−777 (1987). | PubMed | ChemPort |
8. Yang, C. C. & Nash, H. Cell 57, 869−880 (1989). | Article | PubMed | ISI | ChemPort |
9. Schultz, S. C., Shields, G. C. & Steitz, T. A. Science 253, 1001−1007 (1991). | PubMed | ISI | ChemPort |
10. Wilson, G. G. Gene 74, 281−289 (1988). | Article | PubMed | ChemPort |
11. Valentin-Hansen, P. et al. Molec. Microbiol. 3, 1385−1390 (1989). | ChemPort |
12. Bird, A. P. & Southern, E. M. J. molec. Biol. 118, 27−47 (1978). | Article | PubMed | ISI | ChemPort |
13. Yamada, M. & Saier, M. J. biol. Chem. 262, 5455−5462 (1987). | PubMed | ChemPort |
14. Lengeler, J. & Steinberger, H. Molec. gen. Genet. 164, 163−170 (1978). | Article | PubMed | ChemPort |
15. Sabounn, D. & Beckwith, J. J. Bact. 122, 338−340 (1975). | PubMed |
16. Piette, J. et al. Proc. natn. Acad. Sci. U.S.A. 81, 4134−4138 (1984). | ChemPort |
17. Lyons, S. M. & Schendel, P. F. J. Bact. 159, 421−423 (1984). | PubMed | ChemPort |
18. Campbell, J. L. & Kleckner, N. Cell 62, 967−979 (1990). | Article | PubMed | ISI | ChemPort |
19. Ephrussi, A., Church, G. M., Tonegawa, S. & Gilbert, W. Science 227, 134−140 (1985). | PubMed | ISI | ChemPort |
20. Becker, M. M. & Wang, J. C. Nature 309, 682−687 (1984). | Article | PubMed | ISI | ChemPort |
21. Cartwright, I. & Kelly, S. E. Bio Techniques 11, 188−203 (1991). | ChemPort |
22. Saluz, H. P., Wiebauer, K. & Wallace, A. Trends Genet. 7, 207−211 (1991). | PubMed | ChemPort |
23. Singh, J. & Klar, A. J. S. Genes Dev. 6, 186−196 (1992). | PubMed | ISI | ChemPort |
24. Feher, Z., Schlagman, S. L. Miner, Z. & Hattman, S. Curr. Genet. 16, 461−464 (1989). | PubMed | ChemPort |
25. Kwoh, T. J. et al. Nucleic Acids Res. 16, 11489−11505 (1988). | PubMed | ChemPort |
26. Vovis, G. F. & Lacks, S. J. J. molec. Biol 115, 525−538 (1977). | Article | PubMed | ISI | ChemPort |
27. Gribskov, M., Luthy, R., & Eisenberg, D. Meth. Enzym. 183, 146−159 (1990). | PubMed | ChemPort |
28. Stormo, G. D. & Hartzell, G. W. Proc. natn. Acad. Sci. U.S.A. 86, 1183−1187 (1989). | ChemPort |
29. Jiang, W. et al. Molec. Microbiol. 4, 2003−2006 (1990). | ChemPort |
30. Davis, T., Yamada, M., Elgort, M. & Saier, M. H. Molec. Microbiol. 2, 405−412 (1988). | ChemPort |
31. Silverman, M. & Simon, M. J. Bact. 120, 1196−1203 (1974). | PubMed | ChemPort |
32. Bartlett, B. H., Frantz, B. B. & Matsumura, P. J. Bact. 170, 1575−1581 (1988). | PubMed |
33. Brissette, J. L., Weiner, L., Ripmaster, T. L. & Model, P. Genbank 69.0 (1991).
34. Shea, C. M. & Mclntosh, M. A. Genbank 69.0 (1991).
35. E'oright, R. in Molecular Structure and Biological Activity (eds Griffen, J. & Duax, W.) (Elsevier Scientific, New York, 1982).
36. Gunasekera, A., Ebright, Y. W. & Ebright, R. H. Nucleic Acids Res. 18, 6853−6856 (1990). | PubMed | ChemPort |
37. Church, G. M. & Gilbert, W. Proc. natn. Acad. Sci. U.S.A. 81, 1991−1995 (1984). | ChemPort |
38. Church, G. M. & Kieffer-Higgins, S. Science 240, 185−188 (1988). | PubMed | ISI | ChemPort |
  © 1992 Nature Publishing Group

 

 

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