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Formation of a nucleoprotein complex containing Tn7 and its target DNA regulates transposition initiation by Zachary Skelding, Robert Sarnovsky, and Nancy.

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Presentation on theme: "Formation of a nucleoprotein complex containing Tn7 and its target DNA regulates transposition initiation by Zachary Skelding, Robert Sarnovsky, and Nancy."— Presentation transcript:

1 Formation of a nucleoprotein complex containing Tn7 and its target DNA regulates transposition initiation by Zachary Skelding, Robert Sarnovsky, and Nancy L. Craig EMBO J. Volume 21(13):3494-3504 July 1, 2002 ©2002 by European Molecular Biology Organization

2 (A) Schematic of Tn7 higher ‐ order nucleoprotein complexes. Zachary Skelding et al. EMBO J. 2002;21:3494-3504 ©2002 by European Molecular Biology Organization

3 (A) The TnsABC+D (L,R)–Tgt complex contains target DNA and requires a donor DNA with two Tn7 ends. Zachary Skelding et al. EMBO J. 2002;21:3494-3504 ©2002 by European Molecular Biology Organization

4 The TnsABC+D (L,R)–Tgt complex is competent for transposition. Zachary Skelding et al. EMBO J. 2002;21:3494-3504 ©2002 by European Molecular Biology Organization

5 Mutagenesis of the C ‐ terminus of TnsB strongly decreases transposition and interaction with the target DNA. (A) Alignment of the amino acid sequences from the C ‐ terminus of Tn7 TnsB with Tn5468 TnsB. Zachary Skelding et al. EMBO J. 2002;21:3494-3504 ©2002 by European Molecular Biology Organization

6 TnsB and TnsCA225V can mediate formation of a (L,R)–Tgt complex, and this reaction depends on residues within the C ‐ terminus of TnsB. Zachary Skelding et al. EMBO J. 2002;21:3494-3504 ©2002 by European Molecular Biology Organization

7 The TnsABC+D (L,R)–Tgt complex is more stable than the TnsBC+D (L,R)–Tgt complex. Zachary Skelding et al. EMBO J. 2002;21:3494-3504 ©2002 by European Molecular Biology Organization

8 Mutation of the C ‐ terminus of TnsB does not significantly affect TnsC ‐ stimulated TnsAB cleavage. Zachary Skelding et al. EMBO J. 2002;21:3494-3504 ©2002 by European Molecular Biology Organization


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