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. 2003 Jan 31;32(4):239–248. doi: 10.1046/j.1365-2184.1999.3240239.x

Cell cycle‐dependent regulation of the DNA‐dependent protein kinase

A Nilsson 1, F Sirzén 1, R Lewensohn 1, N Wang 1, S Skog 1
PMCID: PMC6726331  PMID: 10614713

Abstract

Abstract. Human DNA‐dependent protein kinase (DNA‐PK) is a nuclear‐localized serine/threonine protein kinase. The holoenzyme consists of a catalytic subunit with a molecular mass of 465 kDa and a DNA‐binding heterodimer Ku86/70. The kinase has been implicated in a variety of nuclear processes including V(D)J recombination, double‐strand break repair, and transcription. Cells with defective DNA‐PK activity show increased radiosensitivity and lack of V(D)J recombination. To study DNA‐PK activity during the cell cycle, HeLa cells were separated by elutriation centrifugation into different cell cycle compartments based on cellular size. DNA‐PK activity was found to vary during the cell cycle. The kinase activity was lowest during G1 phase and increased dramatically as the cells entered S phase and remained high during the G2‐phase. The subcellular distribution of DNA‐PKcs is relocalized from the cytoplasm during M and G1 phases to the nucleus during G1–S phase transition and S phase. Expression of both the catalytic subunit and the Ku86/70 heterodimer was found to be constant throughout the cell cycle. This study demonstrates that DNA‐PK activity as well as its subcellular localization fluctuates during the cell cycle. In addition, the distribution of DNA‐PK during M phase corresponds with low DNA‐PK activity.

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