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Lecture 11

BIO372 - Lecture 11.pdf

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Gordon Anderson

Lecture 11 February-25-14 10:14 AM Recombination:nicking of DNA, single stranded overhang attacks the duplex, strand displacement, further synthesis of DNA, intermediatejunction E. coli RecBCD Pathway of Homologous Recombination I SometimesRecABCD -recA helps in homologysearch Repair is the initial condition that drove evolution BCD - encodes for different subunits Under conditions of high magnesium, the activity is higher Acts as endonuclease,and degrades and unwinds DNA, until it reaches a Chi site (a consensus). Chi sites serve as initiator for replication. Overhang gets coded by recA protein. 30 bp is not enough, if over this, the D-loop will form (displacement-formation),after which recA is released ATP is needed to unload (no ATP no recombination!) HOLLIDAY junction - the polynucleotideswitch between duplex - They are heteroduplex which contain potential mismatches(contain both blue and red from different parents) - Holliday junction migration - A diffusion process, it is neutral, slow process, less enzymes are - Holliday junction migration - A diffusion process, it is neutral, slow process, less enzymes are involved G and H resolutions are two possible resolutions - equally likely to happen RuvC - enzyme that drives the resolution H is the parental resolution (heteroduplexes: non-crossover recombinants)- blue with blue, red with red G is another resolution (crossoverrecombinants) - red with blue Binding of RecA to single-stranded DNA Single stranded areas are much thicker than double stranded areas - protein are bound to DNA (they are huge) At the right pic, same thing, the single stranded DNA are fat The left pic is a nicer experiment than the right one because there's no control
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