Comptes Rendus
no. 5
Biophysique à l'échelle de la molécule unique/Single molecule biophysics
Mechanical opening of DNA by micro manipulation and force measurements
[Ouverture mécanique de la molécule d'ADN par micro-manipulation et mesure de force]
Ulrich Bockelmann1  ; B. Essevaz-Roulet1  ; Philippe Thomen1  ; François Heslot1
1 Laboratoire de physique de la matière condensée, École normale supérieure, 24, rue Lhomond, 75005 Paris, France
Comptes Rendus. Physique, Volume 3 (2002) no. 5, pp. 585-594.

Dans cet article nous résumons une partie de notre travail sur l'ouverture mécanique de l'ADN. Nous avons préparé des constructions moléculaires qui nous permettent d'attacher les deux brins complémentaires d'une extrémité d'une molécule unique d'ADN du bactériophage λ, séparément à une lamelle de microscope et à une bille microscopique. Dans une première série d'expériences, un levier souple de verre, agissant comme capteur de force, est attaché à la bille et sa deflexion est mesurée avec un microscope optique. Dans une seconde série, nous utilisons un piège optique interférométrique pour capturer la bille et mesurer son déplacement avec une résolution nanométrique. L'échantillon est déplacé lentement par rapport au système de mesure de force, induisant une ouverture progressive de la double hélice. La force, mesurée pendant cette ouverture mécanique, montre une variation caractéristique qui est reliée à la séquence des paires de base de la molécule d'ADN.

In this paper we summarize part of our work on the mechanical unzipping of DNA. We have prepared molecular constructions which allow us to attach the two complementary strands of one end of a single DNA molecule of the bacteriophage λ separately to a glass microscope slide and a microscopic bead. In a first series of experiments, a soft microneedle acting as a force sensor is attached to the bead and its deflection is measured with an optical microscope. In a second series, we use an optical trapping interferometer to capture the bead and to measure its displacement to nm resolution. The sample is slowly displaced with respect to the force measurement device, leading to a progressive opening of the double helix. The force measured during this mechanical opening shows a characteristic variation which is related to the base pair sequence of the DNA molecule.

Reçu le :
Accepté le :
Publié le :
DOI : 10.1016/S1631-0705(02)01342-7
Keywords: micromanipulation, optical tweezers, DNA unzipping
Mot clés : micromanipulation, pinces optiques, ouverture de l'ADN
Ulrich Bockelmann 1 ; B. Essevaz-Roulet 1 ; Philippe Thomen 1 ; François Heslot 1

1 Laboratoire de physique de la matière condensée, École normale supérieure, 24, rue Lhomond, 75005 Paris, France 
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Ulrich Bockelmann; B. Essevaz-Roulet; Philippe Thomen; François Heslot. Mechanical opening of DNA by micro manipulation and force measurements. Comptes Rendus. Physique, Volume 3 (2002) no. 5, pp. 585-594. doi : 10.1016/S1631-0705(02)01342-7. https://comptes-rendus.academie-sciences.fr/physique/articles/10.1016/S1631-0705(02)01342-7/

[1] S.B. Smith; L. Finzi; C. Bustamante Direct mechanical measurements of the elasticity of single DNA molecules by using magnetic beads, Science, Volume 258 (1992), pp. 1122-1126

[2] G.U. Lee; L.A. Chrisey; R.J. Colton Direct measurement of the forces between complementary strands of DNA, Science, Volume 266 (1994), pp. 771-773

[3] Ph. Cluzel; A. Lebrun; C. Heller; R. Lavery; J.-L. Viovy; D. Chatenay; F. Caron DNA: an extensible molecule, Science, Volume 271 (1996), pp. 792-794

[4] S.B. Smith; Y. Cui; C. Bustamante Overstretching B-DNA: the elastic response of individual double-stranded and single-stranded DNA molecules, Science, Volume 271 (1996), pp. 795-799

[5] M.D. Wang; H. Yin; R. Landick; J. Gelles; S.M. Block Stretching DNA with optical tweezers, Biophys. J., Volume 72 (1997), pp. 1335-1346

[6] B. Essevaz-Roulet; U. Bockelmann; F. Heslot Mechanical separation of the complementary strands of DNA, Proc. Natl. Acad. Sci. USA, Volume 94 (1997), pp. 11935-11940

[7] U. Bockelmann; B. Essevaz-Roulet; F. Heslot Molecular stick–slip motion revealed by opening DNA with piconewton forces, Phys. Rev. Lett., Volume 79 (1997), pp. 4489-4492

[8] U. Bockelmann; B. Essevaz-Roulet; F. Heslot DNA strand separation studied by single molecule force measurement, Phys. Rev. E, Volume 58 (1998), pp. 2386-2394

[9] J.F. Allemand; D. Bensimon; R. Lavery; V. Croquette Stretched and overwound DNA forms a Pauling-like structure with exposed bases, Proc. Natl. Acad. Sci. USA, Volume 95 (1998), pp. 14152-14157

[10] T.R. Strick; J.F. Allemand; D. Bensimon; A. Bensimon; V. Croquette The elasticity of a single supercoiled DNA molecule, Science, Volume 271 (1996), pp. 1835-1837

[11] J.F. Léger; G. Romano; A. Sarkar; J. Robert; L. Bourdieu; D. Chatenay; J.F. Marko Structural transitions of a twisted and stretched DNA molecule, Phys. Rev. Lett., Volume 83 (1999), pp. 1066-1069

[12] M. Rief; H. Clausen-Schaumann; H.E. Gaub Sequence dependent mechanics of single DNA molecules, Nature Struct. Biol., Volume 6 (1999), pp. 346-349

[13] H. Clausen-Schaumann; M. Rief; C. Tolksdorf; H.E. Gaub Mechanical stability of single DNA molecules, Biophys. J., Volume 78 (2000), pp. 1997-2007

[14] C. Bustamante; S.B. Smith; J. Liphardt; D. Smith Single-molecule studies of DNA mechanics, Curr. Opin. Struct. Biol., Volume 10 (2000), pp. 279-285

[15] U. Bockelmann; Ph. Thomen; B. Essevaz-Roulet; V. Viasnoff; F. Heslot Unzipping DNA with optical tweezers: high sequence sensitivity and force flips, Biophys. J., Volume 82 (2002), p. 1537

[16] Ph. Thomen; U. Bockelmann; F. Heslot Rotational drag on DNA: a single molecule experiment, Phys. Rev. Lett., Volume 88 (2002), p. 248102

[17] S. Cocco; R. Monasson; J.F. Marko Force and kinetic barriers to unzipping of the DNA double helix, Proc. Natl. Acad. Sci. USA, Volume 98 (2001), pp. 8608-8613

[18] D.K. Lubensky; D.R. Nelson Pulling pinned polymers and unzipping DNA, Phys. Rev. Lett., Volume 85 (2000), pp. 1572-1575

[19] P. Nelson Transport of torsional stress in DNA, Proc. Natl. Acad. Sci. USA, Volume 96 (1999), pp. 14342-14347

[20] R.E. Thompson; E.D. Siggia Physical limits on the mechanical measurement of the secondary structure of biomolecules, Europhys. Lett., Volume 31 (1995), pp. 335-340

[21] J.-L. Viovy; Ch. Heller; F. Caron; Ph. Cluzel; D. Chatenay Sequencing of DNA by mechanical opening of the double helix: a theoretical evaluation, C. R. Acad. Sci. Paris, Volume 317 (1994), pp. 795-800

[22] K. Svoboda; S.M. Block Biological applications of optical forces, Annu. Rev. Biophys. Biomol. Struct., Volume 23 (1994), pp. 247-285

[23] W. Denk; W.W. Webb Optical measurements of picometer displacements of transparent microscopic objects, Appl. Opt., Volume 29 (1990), pp. 2382-2391

[24] R.M.A. Azzam; N.M. Bashara Ellipsometry and Polarized Light, North-Holland, Amsterdam, 1989

[25] F. Gittes; C.F. Schmidt Signals and noise in micromechanical measurements, Methods Cell Biol., Volume 55 (1998), pp. 129-156

[26] P. Bowden; D. Tabor Friction and Lubrication of Solids, Clarendon, Oxford, 1950

[27] Ch. Scholz The Mechanics of Earthquakes and Faulting, Cambridge University Press, Cambridge, 1990

[28] T. Baumberger; F. Heslot; B. Perrin Crossover from creep to inertial motion in friction dynamics, Nature, Volume 367 (1994), p. 544

[29] J. Liphardt; B. Onoa; S.B. Smith; I. Tinoco; C. Bustamante Reversible unfolding of single RNA molecules by mechanical force, Science, Volume 292 (2001), pp. 733-737

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