Single-stranded heteroduplex intermediates in lambda Red homologous recombination.

ABSTRACT:

Background

The Red proteins of lambda phage mediate probably the simplest and most efficient homologous recombination reactions yet described. However the mechanism of dsDNA recombination remains undefined.

Results

Here we show that the Red proteins can act via full length single stranded intermediates to establish single stranded heteroduplexes at the replication fork. We created asymmetrically digestible dsDNA substrates by exploiting the fact that Redalpha exonuclease activity requires a 5' phosphorylated end, or is blocked by phosphothioates. Using these substrates, we found that the most efficient configuration for dsDNA recombination occurred when the strand that can prime Okazaki-like synthesis contained both homology regions on the same ssDNA molecule. Furthermore, w

SUBMITTER: Maresca M 

PROVIDER: S-EPMC2918612 | biostudies-literature | 2010 Jul

REPOSITORIES: biostudies-literature