Unknown

Dataset Information

0

DNA structure specificity of Rap endonuclease.


ABSTRACT: The Rap protein of phage lambda is an endonuclease that nicks branched DNA structures. It has been proposed that Rap can nick D-loops formed during phage recombination to generate splice products without the need for the formation of a 4-strand (Holliday) junction. The structure specificity of Rap was investigated using a variety of branched DNA molecules made by annealing partially complementary oligo-nucleotides. On Holliday junctions, Rap endonuclease shows a requirement for magnesium or manganese ions, with Mn(2+)supporting 5-fold more cleavage than Mg(2+). The location of endonuclease incisions was determined on 3'-tailed D-loop, bubble, flayed duplex, 5'-flap and Y junction DNA substrates. In all cases, Rap preferentially cleaves at the branch point of these molecules. With a flayed duplex, incisions are made in the duplex adjacent to the single-strand arms. Comparison of binding and cleavage specificities revealed that Rap is highly structure-specific and exhibits a clear preference for 4- and 3-stranded DNA over Y and flayed duplex DNA. Almost no binding or cleavage was detected with duplex, partial duplex and single-stranded DNA. Thus Rap endonuclease shows a bias for structures that resemble D-loop and Holliday junction recombination intermediates.

SUBMITTER: Sharples GJ 

PROVIDER: S-EPMC148684 | BioStudies | 1999-01-01T00:00:00Z

SECONDARY ACCESSION(S): AF034975

REPOSITORIES: biostudies

Similar Datasets

2013-01-01 | S-EPMC3864405 | BioStudies
2013-01-01 | S-EPMC3834828 | BioStudies
1000-01-01 | S-EPMC2613635 | BioStudies
2011-01-01 | S-EPMC3157427 | BioStudies
1000-01-01 | S-EPMC4678824 | BioStudies
2007-01-01 | S-EPMC2094049 | BioStudies
2013-01-01 | S-EPMC3834835 | BioStudies
1000-01-01 | S-EPMC2587595 | BioStudies
1000-01-01 | S-EPMC2268786 | BioStudies
2001-01-01 | S-EPMC125760 | BioStudies