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Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
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The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...
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Translesion (TLS) polymerases rescue stalled DNA polymerases at sites of damaged bases by replacing the replicative polymerase and installing a nucleotide across the damaged site. Doing so, TLS allows additional time for the cell to repair the damage before resuming regular DNA replication.
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In contrast to the lytic cycle, phages infecting bacteria via the lysogenic cycle do not immediately kill their host cell. Instead, they combine their genome with the host genome, allowing the bacteria to replicate the phage DNA along with the bacterial genome. The incorporated copy of the phage genome is called the prophage. Some prophages can re-activate and enter the lytic cycle. This often occurs in response to a perturbation, such as DNA damage, but can also transpire in the absence of...
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Cells are sometimes infected by more than one virus at once. When two viruses disassemble to expose their genomes for replication in the same cell, similar regions of their genomes can pair together and exchange sequences in a process called recombination. Alternatively, viruses with segmented genomes can swap segments in a process called reassortment.
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DNA replication is initiated at sites containing predefined DNA sequences known as origins of replication. DNA is unwound at these sites by the minichromosome maintenance (MCM) helicase and other factors such as Cdc45 and the associated GINS complex.The unwound single strands are protected by replication protein A (RPA) until DNA polymerase starts synthesizing DNA at the 5’ end of the strand in the same direction as the replication fork. To prevent the replication fork from falling apart,...
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Following Cell-fate in E. coli After Infection by Phage Lambda
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Bacteriophage lambda site-specific recombination.

Gregory D Van Duyne1, Arthur Landy2

  • 1Department of Biochemistry & Biophysics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, USA.

Molecular Microbiology
|February 19, 2024
PubMed
Summary
This summary is machine-generated.

Bacteriophage λ site-specific recombination involves precise DNA integration and excision into bacterial chromosomes. Understanding these pathways reveals intricate regulation mechanisms essential for host cell physiology.

Keywords:
DNA bendingHolliday junctionbacteriophage lambdaintegrationsite‐specific recombinationtyrosine recombinaseviral excisionviral integration

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Area of Science:

  • Molecular Biology
  • Genetics
  • Virology

Background:

  • Bacteriophage lambda (λ) utilizes site-specific recombination for integrating its chromosome into the bacterial host.
  • This process involves complex, regulated DNA manipulation essential for the viral life cycle and host-bacterial interactions.

Purpose of the Study:

  • To review the current understanding of the mechanisms governing bacteriophage λ site-specific recombination.
  • To highlight key studies from the past decade focusing on the regulation and execution of these pathways.

Main Methods:

  • Analysis of protein-DNA complex structures, particularly four-way Holliday junction intermediates.
  • Review of studies investigating the roles of phage-encoded Int and Xis proteins and host-encoded Integration Host Factor (IHF) and Factor for Inversion Stimulation (Fis).

Main Results:

  • Detailed insights into the mechanisms, directionality, and regulation of integrative and excisive recombination pathways.
  • Identification of specific DNA sequences (240 bp phage, 21 bp bacterial) and protein binding sites crucial for recombination.
  • Elucidation of how differential protein utilization dictates pathway directionality.

Conclusions:

  • The intricate regulation of bacteriophage λ recombination is tightly linked to bacterial host physiology.
  • Structural studies of recombination intermediates have significantly advanced the understanding of this complex biological process.