Sequence-directed DNA translocation by purified FtsK

Science

Volumn 307, Issue 5709, 2005, Pages 586-590

  Pease, Paul J ^a   Levy, Oren ^a   Cost, Gregory J ^a   Gore, Jeff ^a   Piacin, Jerod L ^a   Sherratt, David ^b   Bustamante, Carlos ^a   Cozzarelli, Nicholas R ^a  

^a UNIVERSITY OF CALIFORNIA   (United States)

^b UNIVERSITY OF OXFORD   (United Kingdom)

Author keywords

[No Author keywords available]

Indexed keywords

ADENOSINETRIPHOSPHATE; CHROMOSOMES; DNA SEQUENCES; ESCHERICHIA COLI; MOLECULAR STRUCTURE; PROTEINS;

BIDIRECTIONAL MOTORS; DNA TRANSLOCATION; MOLECULAR MOTORS;

DNA;

ADENOSINE TRIPHOSPHATE; CARRIER PROTEIN; PROTEIN FTSK; UNCLASSIFIED DRUG;

DNA;

ARTICLE; BACTERIAL CHROMOSOME; DISSOCIATION; DNA SEQUENCE; ENERGY RESOURCE; ENZYME PURIFICATION; ENZYME SUBSTRATE; ESCHERICHIA COLI; NONHUMAN; PRIORITY JOURNAL; PROTEIN BINDING; PROTEIN DNA INTERACTION;

ALGORITHMS; BACTERIOPHAGE LAMBDA; BASE SEQUENCE; CHROMOSOMES, BACTERIAL; DNA, BACTERIAL; DNA, SUPERHELICAL; DNA, VIRAL; ESCHERICHIA COLI; ESCHERICHIA COLI PROTEINS; KINETICS; MEMBRANE PROTEINS; MODELS, BIOLOGICAL; MOLECULAR MOTOR PROTEINS; NUCLEIC ACID CONFORMATION;

EID: 12844282326     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.1104885     Document Type: Article

References (34)

1. M. R. Singleton, D. B. Wigley, EMBO J. 22, 4579 (2003).
2. J. Errington, J. Bath, L. J. Wu, Nature Rev. Mol. Cell Biol. 2, 538 (2001).
3. G. D. Recchia, M. Aroyo, D. Wolf, G. Blakely, D. J. Sherratt, EMBO J. 18, 5724 (1999).
4. F. X. Barre et al., Proc. Natl. Acad. Sci. U.S.A. 98, 8189 (2001).
5. E. C. Hendricks, H. Szerlong, T. Hill, P. Kuempel, Mol. Microbiol. 36, 973 (2000).
6. T. L. Raoul Tan, R. Kanaar, C. Wyman, DNA Repair (Amst.) 2, 787 (2003).
7. G. Liu, C. C. Draper, W. D. Donachie, Mol. Microbiol. 29, 893 (1998).
8. G. Blakely, S. Colloms, G. May, M. Burke, D. J. Sherratt, New Biol. 3, 789 (1991).
9. J. Corre, J. M. Louarn, J. Bacteriol. 184, 3801 (2002).
10. I. F. Lau et al., Mol. Microbiol. 49, 731 (2003).
11. O. Espeli, C. Lee, K. J. Marians, J. Biol. Chem. 278, 44639 (2003).
12. Materials and methods are available as supporting material on Science Online.
13. L. Aussel et al., Cell 108, 195 (2002).
14. D. Ristic, C. Wyman, C. Paulusma, R. Kanaar, Proc. Natl. Acad. Sci. U.S.A. 98, 8454 (2001).
15. O. A. Saleh, C. Perals, F. X. Barre, J. F. Allemand, EMBO J. 23, 2430 (2004).
16. M. D. Wang et al., Science 282, 902 (1998).
17. N. R. Forde, D. Izhaky, G. R. Woodcock, G. J. Wuite, C. Bustamante, Proc. Natl. Acad. Sci. U.S.A. 99, 11682 (2002).
18. D. E. Smith et al., Nature 413, 748 (2001).
19. P. R. Bianco et al., Nature 409, 374 (2001).
20. G. J. Wuite, S. B. Smith, M. Young, D. Keller, C. Bustamante, Nature 404, 103 (2000).
21. We use the term "motor" to mean an enzyme that translocates or binds DNA at one point, with a molecular structure yet to be established.
22. R. Wessel, J. Schweizer, H. Stahl, J. Virol. 66, 804 (1992).
23. FtsK stopped translocating above ∼63 pN, presumably because of DNA distortion, but remained bound and continued translocating if the force was lowered below the over-stretch transition.
24. S. B. Smith, Y. Cui, C. Bustamante, Science 271, 795 (1996).
25. This experiment also supports our conclusion that we are observing the activity of a single FtsK complex. If multiple complexes were bound to the DNA, we would expect to see by chance occasional reeling in from above and below the observed particle.
26. It is difficult to measure a conventional stall force, because in almost all cases, the DNA loop releases before the motor velocity falls to zero.
27. Because processive motion is observed against a force of ∼63 pN, we can place an upper limit of 1.6 nm (100 pN nm/63 pN) on the step size of the motor. This upper bound was calculated by assuming that a single ATP molecule is hydrolyzed per step and by recognizing that the efficiency must be less than 100%.
28. D. L. Kaplan, M. O'Donnell, Mol. Cell 15, 453 (2004).
29. S. L. Salzberg, A. J. Salzberg, A. R. Kerlavage, J. F. Tomb, Gene 217, 57 (1998).
30. M. Spies et al., Cell 114, 647 (2003).
31. J. Bath, L. J. Wu, J. Errington, J. C. Wang, Science 290, 995 (2000).
32. L. Postow, C. D. Hardy, J. Arsuaga, N. R. Cozzarelli, Genes Dev. 18, 1766 (2004).
33. J. Louarn, F. Cornet, V. Francois, J. Patte, J. M. Louarn, J. Bacteriol. 176, 7524 (1994).
34. 50C Histagged construct and S. B. Smith for expert assistance. This work was supported by NIH grants GM31657 (N.R.C.), GM32543 (C.B.), GM07232-27 (J.L.P.), Wellcome Trust and the Royal Society (D.S.), Ruth L. Kirschstein National Research Service Award, grant GM08295-15 (P.J.P.), Fannie and John Hertz Foundation (J.G.), Damon Runyon Cancer Research Foundation grant 1702-02 (G.J.C.), and U.S. Department of Energy grants DE-AC03-76DF00098, GTL2BN "Microscopies of MolecularMachines," and SNANOB "Design of Autonomous Nanobots" (C.B.).