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DNA Recognition in Procaryotes by Helix-Turn-Helix Motifs.

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Presentation on theme: "DNA Recognition in Procaryotes by Helix-Turn-Helix Motifs."— Presentation transcript:

1 DNA Recognition in Procaryotes by Helix-Turn-Helix Motifs

2 Lysogeny vs. Lysis

3 Lysogenic Lytic

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5 Cro from phage-monomer

6 Cro from phage-dimer

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8 Repressor from phage-monomer

9 Repressor from phage-dimer

10 helix-turn-helix

11 Cro - DNA recognition helices vs. 34 Å

12 Genetics + Structural Biology

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14 Repressor from 434 phage-dimer

15 10.2 Most bacterial repressors are dimers containing  helices that insert into adjacent major grooves of operator DNA Figure 10-13 Copyright (c) by W.H.Freeman and Company

16 434 repressor-DNA binding BeforeAfter

17 434 cro/repressor-DNA binding

18 434 repressor-DNA binding

19 434 repressor-DNA binding- specific interaction

20 434 repressor-DNA binding- Non-specific interaction

21 Protein-DNA interaction in Helix-turn-helix

22 By itself, an operon is on and RNA polymerase can bind to the promoter and transcribe the genes. Copyright © 2002 Pearson Education, Inc., publishing as Benjamin Cummings Fig. 18.20a

23 However, if a repressor protein, a product of a regulatory gene, binds to the operator, it can prevent transcription of the operon’s genes. –Each repressor protein recognizes and binds only to the operator of a certain operon. –Regulatory genes are transcribed at low rates continuously. Copyright © 2002 Pearson Education, Inc., publishing as Benjamin Cummings Fig. 18.20b

24 trp repressor-monomer

25 trp repressor-dimer

26 trp repressor-DNA

27 10.2 Ligand-induced conformational changes alter affinity of many repressors for DNA Figure 10-14 Tryptophan binding induces a conformational change in the trp aporepressor Copyright (c) by W.H.Freeman and Company

28 10.2 DNase I footprinting assays identify protein-DNA interactions Figure 10-6 Copyright (c) by W.H.Freeman and Company

29 10.2 Gel-shift assays identify protein- DNA interactions Figure 10-7 Copyright (c) by W.H.Freeman and Company

30 10.2 The footprint of RNA polymerase and lac repressor on the lac control region Figure 10-8 Copyright (c) by W.H.Freeman and Company

31 10.2 The lac control region contains three critical cis-acting sites Figure 10-9 Copyright (c) by W.H.Freeman and Company

32 10.2 Positive control of the lac operon is exerted by cAMP-CAP Figure 10-16 CAP = catabolite activator protein Copyright (c) by W.H.Freeman and Company

33 lac repressor-monomer

34 lac repressor(tetramer)-DNA

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36 CAP-DNA

37 10.2 A space-filling model of cAMP- CAP bound to lac promoter DNA Figure 10-18 Copyright (c) by W.H.Freeman and Company

38 10.2 Cooperative binding of cAMP-CAP and RNA polymerase to the lac contol region activates transcription Figure 10-17 Copyright (c) by W.H.Freeman and Company

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46 http://www.biochem.ucl.ac.uk/bsm/prot_dna/prot_dna.html

47 http://www.biochem.ucl.ac.uk/bsm/DNA/server/


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