Loss of homologous recombination or non-homologous end-joining leads to radial formation following DNA interstrand crosslink damage

A. E. Hanlon Newell, A. Hemphill, Y. M N Akkari, J. Hejna, Robb Moses, Susan Olson

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

High levels of interstrand cross-link damage in mammalian cells cause chromatid breaks and radial formations recognizable by cytogenetic examination. The mechanism of radial formation observed following DNA damage has yet to be determined. Due to recent findings linking homologous recombination and non-homologous end-joining to the action of the Fanconi anemia pathway, we speculated that radials might be the result of defects in either of the pathways of DNA repair. To test this hypothesis, we have investigated the role of homologous recombination proteins RAD51 and RAD52, non-homologous end-joining proteins Ku70 and LIG4, and protein MRE11 in radial formation and cell survival following interstrand crosslink damage with mitomycin C. For the studies we used small inhibitory RNA to deplete the proteins from cells, allowing for evaluation of radial formation and cell survival. In transformed normal human fibroblasts, depletion of these proteins increased interstrand crosslink sensitivity as manifested by decreased cell survival and increased radial formation. These results demonstrate that inactivation of proteins from either of the two separate DNA repair pathways increases cellular sensitivity to interstrand crosslinks, indicating each pathway plays a role in the normal response to interstrand crosslink damage. We can also conclude that homologous recombination or non-homologous end-joining are not required for radial formation, since radials occur with depletion of these pathways.

Original languageEnglish (US)
Pages (from-to)174-180
Number of pages7
JournalCytogenetic and Genome Research
Volume121
Issue number3-4
DOIs
StatePublished - Aug 2008

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Homologous Recombination
DNA
Cell Survival
Proteins
DNA Repair
Rad51 Recombinase
Fanconi Anemia
Chromatids
Mitomycin
Cytogenetics
DNA Damage
Fibroblasts
RNA

ASJC Scopus subject areas

  • Genetics
  • Cell Biology

Cite this

Loss of homologous recombination or non-homologous end-joining leads to radial formation following DNA interstrand crosslink damage. / Hanlon Newell, A. E.; Hemphill, A.; Akkari, Y. M N; Hejna, J.; Moses, Robb; Olson, Susan.

In: Cytogenetic and Genome Research, Vol. 121, No. 3-4, 08.2008, p. 174-180.

Research output: Contribution to journalArticle

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