Related Experiment Video

Updated: Jul 24, 2026

Preparation of the Mgm101 Recombination Protein by MBP-based Tagging Strategy
11:40

Preparation of the Mgm101 Recombination Protein by MBP-based Tagging Strategy

Published on: June 25, 2013

Involvement of minor structural proteins in recombination of polyoma virus DNA

S Charbonneau1, D Gendron, E Samson

  • 1Department of Microbiology and Infectious Diseases, The Medical School, Sherbrooke, Québec, J1H 5N4, Canada.

Virology
|December 9, 2000
PubMed

Insights

Mutations in the VP2/VP3 coding sequence of polyoma virus (PV) hinder DNA recombination, suggesting VP2 plays a key role in this process. This impacts PV DNA generation in transfected cells.

Area of Science:

  • Molecular Biology
  • Virology
  • Genetics

Background:

  • Polyoma-mouse chimeric DNA (RmI) with repeated viral sequences facilitates intramolecular recombination.
  • This recombination converts RmI into unit-length polyoma DNA in permissive mouse cells.
  • The process is linked to VmP1, a protein with polyoma VP1 N-terminus and murine C-terminus.

Purpose of the Study:

  • To investigate the role of VP2/VP3 coding sequences in the recombination of polyoma-mouse chimeric DNA (RmI).
  • To determine the impact of mutations in VP2/VP3 on polyoma DNA generation and replication.

Main Methods:

  • Transfection of permissive mouse cells with RmI constructs.
  • Introduction of mutations within the VP2/VP3 coding sequence of RmI.
  • Analysis of polyoma DNA generation, replication, and accumulation post-transfection.

Main Results:

  • Mutations in the VP2/VP3 coding sequence significantly reduced RmI's ability to generate polyoma DNA.
  • These mutations had minimal impact on polyoma DNA replication or accumulation.
  • A mutation affecting VP2 alone was as effective as one affecting both VP2 and VP3.

Conclusions:

  • The VP2/VP3 coding sequence, particularly VP2, is critical for the intramolecular recombination of polyoma-mouse chimeric DNA.
  • VP2 plays a crucial role in polyoma DNA generation, independent of its role in DNA replication or accumulation.

More Related Videos

Purification of Viral DNA for the Identification of Associated Viral and Cellular Proteins08:26

Purification of Viral DNA for the Identification of Associated Viral and Cellular Proteins

Published on: August 31, 2017

Electrophoretic Analysis of Replication Through Structure-Prone DNA Repeats Within the SV40-Based Human Episome05:22

Electrophoretic Analysis of Replication Through Structure-Prone DNA Repeats Within the SV40-Based Human Episome

Published on: September 13, 2024

Related Concept Videos

Viral Recombination00:57

Viral Recombination

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.
The Replisome03:01

The Replisome

DNA replication is carried out by a large complex of proteins that act in a coordinated matter to achieve high-fidelity DNA replication. Together this complex is known as the DNA replication machinery or the replisome.
The synthesis of the leading and lagging strands is a highly coordinated process. To explain this, the “Trombone model” was proposed by Bruce Alberts in 1980. The DNA loop formation starts when a primer is synthesized on the parent lagging strand. The loop grows with the...
Homologous Recombination02:31

Homologous Recombination

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...
Restarting Stalled Replication Forks02:37

Restarting Stalled Replication Forks

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, a...
Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

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.
The recognition sites for Cre recombinase called LoxP...
Homologous Recombination02:31

Homologous Recombination

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...