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Related Experiment Videos

Complementation analysis of the murine scid cell line

M Z Zdzienicka1, W Jongmans, M Oshimura

  • 1Department of Radiation Genetics and Chemical Mutagenesis, University of Leiden, The Netherlands.

Radiation Research
|September 1, 1995
PubMed
Summary

Chinese hamster cell mutants and murine scid cells share defects in DNA double-strand break repair and V(D)J recombination. These studies suggest the V-3 mutant and scid cells are defective in the same gene on human chromosome 8.

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Area of Science:

  • Genetics
  • Molecular Biology
  • Cell Biology

Background:

  • Several X-ray-sensitive Chinese hamster cell mutants exhibit defects in DNA double-strand break (DSB) repair and V(D)J recombination.
  • These mutants fall into three complementation groups: xrs series, XR-1, and V-3.
  • The murine severe combined immunodeficiency (scid) cell line shares this phenotype, prompting investigation into its genetic relationship with the hamster mutants.

Purpose of the Study:

  • To determine if the murine scid mutation represents a new complementation group or belongs to an existing one.
  • To investigate the genetic locus of the defect in V-3 and scid cells.

Main Methods:

  • Cell fusion experiments were conducted between scid cells and hamster mutants from the three complementation groups.

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  • Microcell-mediated chromosome transfer was used to introduce human chromosome 8 into V-3 cells.
  • X-ray sensitivity was assessed in hybrid cell lines to evaluate complementation.

    • Main Results:

    • Fusion of V-3 and scid cells resulted in partial complementation of X-ray sensitivity, suggesting a defect in the same gene.
    • Introduction of human chromosome 8 into V-3 cells partially complemented X-ray sensitivity in most hybrid clones.
    • Introduction of human chromosome 8 into scid cells conferred X-ray resistance in the majority of hybrid clones.

    Conclusions:

    • The V-3 hamster mutant and the scid mutation are defective in the same gene.
    • The defective gene in both V-3 and scid cells is located on human chromosome 8.
    • These findings support the genetic linkage between V-3 and scid mutations.