Over-expression of RAD51 or RAD54 but not RAD51/4 enhances extra-chromosomal homologous recombination in the human sarcoma (HT-1080) cell line

J Biotechnol. 2011 Jun 10;154(1):21-4. doi: 10.1016/j.jbiotec.2011.03.023. Epub 2011 Apr 8.

Abstract

RAD51 and RAD54, members of the RAD52 epistasis group, play key roles in homologous recombination (HR). The efficiency of homologous recombination (HR) can be increased by over-expression of either of them. A vector that allows co-expression of RAD51 and RAD54 was constructed to investigate interactions between the two proteins during extra-chromosomal HR. The efficiency of extra-chromosomal HR evaluated by GFP extra-chromosomal HR was enhanced (110-245%) in different transfected Human sarcoma (HT-1080) cell colonies. We observed that RAD51 clearly promotes extra-chromosomal HR; however, the actions of RAD54 in extra-chromosomal HR were weak. Our data suggest that RAD51 may function as a universal factor during HR, whereas RAD54 mainly functions in other types of HR (gene targeting or intra-chromosomal HR), which involves interaction with chromosomal DNA.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Blotting, Western
  • Cell Line, Tumor
  • Chromosomes, Human / genetics*
  • DNA Helicases
  • DNA-Binding Proteins
  • Flow Cytometry
  • Gene Expression*
  • Genetic Vectors / genetics
  • Humans
  • Nuclear Proteins / genetics*
  • Rad51 Recombinase / genetics*
  • Recombination, Genetic*
  • Sarcoma / genetics*
  • Transgenes / genetics

Substances

  • DNA-Binding Proteins
  • Nuclear Proteins
  • RAD51 protein, human
  • Rad51 Recombinase
  • DNA Helicases
  • RAD54L protein, human