Radiation-induced delayed cell death in a hypomorphic Artemis cell line

Hum Mol Genet. 2006 Apr 15;15(8):1303-11. doi: 10.1093/hmg/ddl050. Epub 2006 Mar 15.

Abstract

Null mutations in Artemis confer a condition described as RS-SCID, in which patients display radiosensitivity combined with severe combined immunodeficiency. Here, we characterize the defect in Artemis in a patient who displayed progressive combined immunodeficiency (CID) and elevated lymphocyte apoptosis. The patient is a compound heterozygote with novel mutations in both alleles, resulting in Artemis proteins with either L70 deletion or G126D substitution. Both mutational changes impact upon Artemis function and a fibroblast cell line derived from the patient (F96-224) has greatly reduced Artemis protein. In contrast to Artemis null cell lines, which fail to repair a subset of DNA double strand breaks (DSBs) induced by ionizing radiation, F96-224 cells show slow but residual DSB rejoining. Despite showing intermediate cellular and clinical features, F96-224 cells are as radiosensitive as Artemis null cell lines. We developed a FACS-based assay to examine cell division and cellular characteristics for 10 days following exposure to ionizing radiation (2 and 4 Gy). This analysis demonstrated that F96-224 cells show delayed cell death when compared with rapid growth arrest of an Artemis null cell line, and the emergence of a cycling population shown by a control line. F96-224 cells also display elevated chromosome aberrations when compared with control cells. F96-224 therefore represents a novel phenotype for a hypomorphic cell line. We suggest that delayed cell death contributes to the progressive CID phenotype of the Artemis patient.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Death / radiation effects*
  • Cell Division / drug effects
  • Cell Line
  • DNA Damage / radiation effects
  • DNA Repair
  • DNA-Binding Proteins
  • Endonucleases
  • Fibroblasts / cytology
  • Fibroblasts / metabolism
  • Heterozygote
  • Humans
  • Kinetics
  • Mutation
  • Nuclear Proteins / genetics
  • Nuclear Proteins / metabolism*
  • Severe Combined Immunodeficiency / metabolism
  • VDJ Recombinases / metabolism

Substances

  • DNA-Binding Proteins
  • Nuclear Proteins
  • VDJ Recombinases
  • DCLRE1C protein, human
  • Endonucleases