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27 and 29 September, 2006 Chapter 11 Transposition and Site-Specific Recombination

27 and 29 September, 2006 Chapter 11 Transposition and Site- Specific Recombination

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Page 1: 27 and 29 September, 2006 Chapter 11 Transposition and Site- Specific Recombination

27 and 29 September, 2006

Chapter 11

Transposition and Site-Specific Recombination

Page 2: 27 and 29 September, 2006 Chapter 11 Transposition and Site- Specific Recombination

Overview• Conservative Site-Specific Recombination (CSSR) may involve

insertion, deletion, or inversion of DNA sequences.• Site-specific recombinases have a mechanism that includes a DNA-

protein covalent intermediate.• CSSR may be regulated by the presence or absence of accessory

proteins.• Resolvases are CSSR recombinases that disentangle circular

chromosomes.• Transposons move using recombination pathways.• Transposons may be autonomous or nonautonomous.• DNA transposons and viral-like retrotransposons move via a cut/paste

mechanism.• Retrotransposon movement involves reverse transcription.• Some transposons regulate copy number or control target site selection

through the use of proteins or antisense RNA.• V(D)J recombination uses regulated, specific recombination to generate

immune diversity.

Page 3: 27 and 29 September, 2006 Chapter 11 Transposition and Site- Specific Recombination

Recombination and Transposition

Page 4: 27 and 29 September, 2006 Chapter 11 Transposition and Site- Specific Recombination

CSSR: Prophage Insertion

Page 5: 27 and 29 September, 2006 Chapter 11 Transposition and Site- Specific Recombination

Three Classes of CSSR

Page 6: 27 and 29 September, 2006 Chapter 11 Transposition and Site- Specific Recombination

Recombinase Recognition Sites

Page 7: 27 and 29 September, 2006 Chapter 11 Transposition and Site- Specific Recombination

Recombinase Mechanism

Page 8: 27 and 29 September, 2006 Chapter 11 Transposition and Site- Specific Recombination

Serine Recombinases

Page 9: 27 and 29 September, 2006 Chapter 11 Transposition and Site- Specific Recombination

Tyrosine Recombinases

Page 10: 27 and 29 September, 2006 Chapter 11 Transposition and Site- Specific Recombination

Mechanism of Cre

Recombinase

Page 11: 27 and 29 September, 2006 Chapter 11 Transposition and Site- Specific Recombination

Cre-DNA Structure

Page 12: 27 and 29 September, 2006 Chapter 11 Transposition and Site- Specific Recombination

Lambda integration

requires architectural

proteins.

Page 13: 27 and 29 September, 2006 Chapter 11 Transposition and Site- Specific Recombination

Int and IHF stabilize bent

DNA.

Page 14: 27 and 29 September, 2006 Chapter 11 Transposition and Site- Specific Recombination

Hin Inversion

Page 15: 27 and 29 September, 2006 Chapter 11 Transposition and Site- Specific Recombination

Hin inversion requires Fis bound at an enhancer.

Page 16: 27 and 29 September, 2006 Chapter 11 Transposition and Site- Specific Recombination

Resolvases disentangle circular DNAs after replication.

Page 17: 27 and 29 September, 2006 Chapter 11 Transposition and Site- Specific Recombination

FtsK Regulation of

the Xer Resolvase

Mechanism

Page 18: 27 and 29 September, 2006 Chapter 11 Transposition and Site- Specific Recombination

FtsK is present at the division closure site.

Page 19: 27 and 29 September, 2006 Chapter 11 Transposition and Site- Specific Recombination

Conservative and Replicative Transposition

Page 20: 27 and 29 September, 2006 Chapter 11 Transposition and Site- Specific Recombination

Transposons in Several Genomes

Page 21: 27 and 29 September, 2006 Chapter 11 Transposition and Site- Specific Recombination

Types of Transposons

Page 22: 27 and 29 September, 2006 Chapter 11 Transposition and Site- Specific Recombination

Cut and Paste Transposition

Page 23: 27 and 29 September, 2006 Chapter 11 Transposition and Site- Specific Recombination

Three Mechanisms for Cleaving the Nontransferred Strand

Page 24: 27 and 29 September, 2006 Chapter 11 Transposition and Site- Specific Recombination

Replicative Transposition

Page 25: 27 and 29 September, 2006 Chapter 11 Transposition and Site- Specific Recombination

Retrotransposon Movement

Page 26: 27 and 29 September, 2006 Chapter 11 Transposition and Site- Specific Recombination

Retrotransposon Movement

Page 27: 27 and 29 September, 2006 Chapter 11 Transposition and Site- Specific Recombination

Retrotransposon Movement

Page 28: 27 and 29 September, 2006 Chapter 11 Transposition and Site- Specific Recombination

DNA Transposases and retroviral integrases are members of the same protein superfamily.

Page 29: 27 and 29 September, 2006 Chapter 11 Transposition and Site- Specific Recombination

LINE poly-A Retrotransposon

Movement

Page 30: 27 and 29 September, 2006 Chapter 11 Transposition and Site- Specific Recombination

Tn10 achieves antisense copy control by overlapping promoters.

Page 31: 27 and 29 September, 2006 Chapter 11 Transposition and Site- Specific Recombination

Antisense Copy

Control

Page 32: 27 and 29 September, 2006 Chapter 11 Transposition and Site- Specific Recombination

Tn10 transposase promoter is active only

when hemimethylated.

Page 33: 27 and 29 September, 2006 Chapter 11 Transposition and Site- Specific Recombination

MuA and MuB participate in selecting Mu transposition

target sequences.

Page 34: 27 and 29 September, 2006 Chapter 11 Transposition and Site- Specific Recombination

MuA disrupts MuB assembly,

conferring transposition

target immunity.

Page 35: 27 and 29 September, 2006 Chapter 11 Transposition and Site- Specific Recombination

Clustered Integration of Yeast Ty Elements

Page 36: 27 and 29 September, 2006 Chapter 11 Transposition and Site- Specific Recombination

Antibody

Page 37: 27 and 29 September, 2006 Chapter 11 Transposition and Site- Specific Recombination

V(D)J Recombination

Page 38: 27 and 29 September, 2006 Chapter 11 Transposition and Site- Specific Recombination

Recombination Signal Sequences

Page 39: 27 and 29 September, 2006 Chapter 11 Transposition and Site- Specific Recombination

V(D)J Recombination

Mechanism

Page 40: 27 and 29 September, 2006 Chapter 11 Transposition and Site- Specific Recombination
Page 41: 27 and 29 September, 2006 Chapter 11 Transposition and Site- Specific Recombination
Page 42: 27 and 29 September, 2006 Chapter 11 Transposition and Site- Specific Recombination
Page 43: 27 and 29 September, 2006 Chapter 11 Transposition and Site- Specific Recombination
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Page 45: 27 and 29 September, 2006 Chapter 11 Transposition and Site- Specific Recombination

Title

Page 46: 27 and 29 September, 2006 Chapter 11 Transposition and Site- Specific Recombination

LINES and SINES Again?