PubMed HealthSearch

PubMed · 6241556

DNA interactions during bacteriophage lambda site-specific recombination.

Abstract

Extensive research on site-specific recombination has provided many details, particularly with respect to the protein-DNA interactions. However, very little is known about the molecular mechanism of recombination during synapsis and strand exchange. Presumably, these steps of recombination involve various forms of DNA-DNA, DNA-protein, and protein-protein interactions. One stage at which DNA-DNA interactions may be occurring is at the level of synapsis where the recombining DNAs are juxtaposed. In this paper we have presented evidence that homology-dependent DNA interactions do occur within the overlap region before strand exchange. This interaction is presumably at the synaptic stage of recombination. Furthermore, we have demonstrated that the homology-dependent interactions require that only one strand of attB have homology to attP. Another stage in recombination at which DNA-DNA interactions could occur is during strand exchange where complementary strands from the recombining parents are paired and resealed. We have also presented evidence that homology-dependent DNA interactions occur during strand exchange prior to the resealing of the strands and that disruption of this interaction results in nonreciprocal recombination. Taken together, these results suggest that DNA-DNA interactions during reciprocal site-specific recombination occur during at least two stages in the reaction.

Explore related subjects

Keep this discovery

Explore connections, maps & timelines

BibTeXRIS

C E Bauer, S D Hesse, J F Gardner, R I Gumport. 1984. DNA interactions during bacteriophage lambda site-specific recombination.. https://doi.org/10.1101/sqb.1984.049.01.079

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related citations

Expression cloning of a zinc-finger cyclic AMP-response-element-binding protein.

In response to specific extracellular signals, intracellular cyclic AMP levels increase, leading to a variety of responses including the alteration of transcription of many eukaryotic genes. This transcriptional effect is frequently mediated through the cyclic AMP-response element (CRE) motif T(T/G)ACGTCA. Using an expression screening approach we have cloned a yeast gene, MSN2, that encodes a 78 kDa protein that recognizes this consensus CRE motif. Phosphorylation of the MSN2 protein by the catalytic subunit of protein kinase A stimulates DNA binding in vitro. Two putative Cys2His2-type zinc fingers present in the C-terminal 79 amino acids of the MSN2 protein are sufficient to confer CRE-binding specificity. Therefore, MSN2 represents a novel CRE-binding protein distinct from the multiple previously characterized basic region-leucine zipper repeat CRE-binding proteins.

Bacteriophage lambda

A hotspot of spontaneous and UV-induced illegitimate recombination during formation of lambda bio transducing phage.

To study the mechanism of spontaneous and UV-induced illegitimate recombination, we examined the formation of the lambda bio specialized transducing phage in Escherichia coli. Because most lambda bio transducing phages have double defects in the red and gam genes and have the capacity to form a plaque on an E. coli P2 lysogen (Spi- phenotype), we selected lambda bio transducing phage by their Spi- phenotype, rather than using the bio marker. We determined sequences of recombination junctions of lambda bio transducing phages isolated with or without UV irradiation and deduced sequences of parental recombination sites. The recombination sites were widely distributed on E. coli bio and lambda DNAs, except for a hotspot which accounts for 57% of UV-induced lambda bio transducing phages and 77% of spontaneously induced lambda bio transducing phages. The hotspot sites on E. coli and lambda DNAs shared a short homology of 9 bp. In addition, we detected direct repeat sequences of 8 bp within and near both the bio and lambda hotspots. A recA mutation did not affect the frequency of the recombination at the hotspot, indicating that this recombination is not a variant of recA-dependent homologous recombination. We discuss a model in which the short homology as well as the direct repeats play essential roles in illegitimate recombination at the hotspot.

Bacteriophage lambda

Efficient large-scale sequencing of the Escherichia coli genome: implementation of a transposon- and PCR-based strategy for the analysis of ordered lambda phage clones.

We have developed a strategy for efficient sequence analysis of the genome of E. coli K-12 using insertions of a Tn5-derived mini-transposon into overlapping ordered lambda phage clones to provide universal primer-binding sites, and PCR amplification of DNA segments adjacent to the insertions. Transposon-containing clones were selected by blue plaque formation on a dnaBamber lacZamber E. coli strain. Insertion points every 0.5-1 kb were identified by 'analytical PCR' and segments between the transposon inserts and phage arms were amplified by 'preparative PCR' using one biotinylated and one non-biotinylated primer. Single strands of amplified DNA fragments were coupled to Streptoavidin-coated paramagnetic beads (Dynabeads M280) through their biotin tails, purified magnetically, and used as templates for fluorescence-based automatic nucleotide sequencing.

Bacteriophage lambda