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1 August 2001 Analysis of Gene Transcription in Cells Lacking DNA-PK Activity
Fredrik Bryntesson, Jenny C. Regan, Penny A. Jeggo, Guillermo E. Taccioli, Michael Hubank
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Abstract

Bryntesson, F., Regan, J. C., Jeggo, P. A., Taccioli, G. E. and Hubank, M. Analysis of Gene Transcription in Cells Lacking DNA-PK Activity. Radiat. Res. 156, 167–176 (2001).

The DNA-dependent protein kinase (DNA-PK), comprised of the Ku70/Ku80 (now known as G22p1/Xrcc5) heterodimer and the catalytic subunit DNA-PKcs (now known as Prkdc), is required for the nonhomologous end joining (NHEJ) pathway of DNA double-strand break repair. The mechanism of action of DNA-PK remains unclear. We have investigated whether DNA-PK regulates gene transcription in vivo after DNA damage using the subtractive hybridization technique of cDNA representational difference analysis (cDNA RDA). Differential transcription, both radiation-dependent and independent, was detected and confirmed in primary mouse embryo fibroblasts from DNA-PKcs–/– and DNA-PKcs / mice. We present evidence that transcription of the extracellular matrix gene laminin alpha 4 (Lama4) is regulated by DNA-PK in a radiation-independent manner. However, screening of both primary and immortalized DNA-PKcs-deficient cell lines demonstrates that the majority of differences were not consistently dependent on DNA-PK status. Similar results were obtained in experiments using KU mutant hamster cell lines, indicating heterogeneity of transcription between closely related cell lines. Our results suggest that while DNA-PK may be involved in limited gene-specific transcription, it does not play a major role in the transcriptional response to DNA damage.

Fredrik Bryntesson, Jenny C. Regan, Penny A. Jeggo, Guillermo E. Taccioli, and Michael Hubank "Analysis of Gene Transcription in Cells Lacking DNA-PK Activity," Radiation Research 156(2), 167-176, (1 August 2001). https://doi.org/10.1667/0033-7587(2001)156[0167:AOGTIC]2.0.CO;2
Received: 6 March 2001; Accepted: 1 May 2001; Published: 1 August 2001
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