Ku70/Ku80 and DNA-dependent protein kinase catalytic subunit modulate RAG-mediated cleavage: Implications for the enforcement of the 12/23 rule

Dennis J. Sawchuk, Jorge Mansilla-Soto, Claudio Alarcon, Netai C. Singha, Hanno Langen, Marco E. Bianchi, Susan P. Lees-Miller, Michel C. Nussenzweig, Patricia Cortes

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

The 12/23 rule is a critical step for regulation of V(D)J recombination. To date, only the RAG proteins and high mobility group protein 1 or 2 have been implicated in 12/23 regulation. Through protein fractionation and biochemical experiments, we find that Ku70/Ku80 and DNA-dependent protein kinase catalytic subunit (DNA-PKcs) modulate RAG-mediated cleavage. Modulation of cleavage by Ku70/80 and DNA-PKcs results in preferential inhibition of 12/12 and 23/23 DNA cleavage, thus increasing 12/23 rule specificity. This observation indicates that DNA repair factors, Ku70/80 and DNA-PKcs, might be present upstream of the DNA cleavage events and not recruited downstream as is currently thought, assigning new nonrepair functions to the DNA-dependent protein kinase.

Original languageEnglish
Pages (from-to)29821-29831
Number of pages11
JournalJournal of Biological Chemistry
Volume279
Issue number28
DOIs
Publication statusPublished - Jul 9 2004

Fingerprint

DNA-Activated Protein Kinase
Catalytic Domain
DNA Cleavage
DNA
V(D)J Recombination
High Mobility Group Proteins
Fractionation
DNA Repair
Proteins
Repair
Modulation
Experiments

ASJC Scopus subject areas

  • Biochemistry

Cite this

Ku70/Ku80 and DNA-dependent protein kinase catalytic subunit modulate RAG-mediated cleavage : Implications for the enforcement of the 12/23 rule. / Sawchuk, Dennis J.; Mansilla-Soto, Jorge; Alarcon, Claudio; Singha, Netai C.; Langen, Hanno; Bianchi, Marco E.; Lees-Miller, Susan P.; Nussenzweig, Michel C.; Cortes, Patricia.

In: Journal of Biological Chemistry, Vol. 279, No. 28, 09.07.2004, p. 29821-29831.

Research output: Contribution to journalArticle

Sawchuk, DJ, Mansilla-Soto, J, Alarcon, C, Singha, NC, Langen, H, Bianchi, ME, Lees-Miller, SP, Nussenzweig, MC & Cortes, P 2004, 'Ku70/Ku80 and DNA-dependent protein kinase catalytic subunit modulate RAG-mediated cleavage: Implications for the enforcement of the 12/23 rule', Journal of Biological Chemistry, vol. 279, no. 28, pp. 29821-29831. https://doi.org/10.1074/jbc.M403706200
Sawchuk, Dennis J. ; Mansilla-Soto, Jorge ; Alarcon, Claudio ; Singha, Netai C. ; Langen, Hanno ; Bianchi, Marco E. ; Lees-Miller, Susan P. ; Nussenzweig, Michel C. ; Cortes, Patricia. / Ku70/Ku80 and DNA-dependent protein kinase catalytic subunit modulate RAG-mediated cleavage : Implications for the enforcement of the 12/23 rule. In: Journal of Biological Chemistry. 2004 ; Vol. 279, No. 28. pp. 29821-29831.
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abstract = "The 12/23 rule is a critical step for regulation of V(D)J recombination. To date, only the RAG proteins and high mobility group protein 1 or 2 have been implicated in 12/23 regulation. Through protein fractionation and biochemical experiments, we find that Ku70/Ku80 and DNA-dependent protein kinase catalytic subunit (DNA-PKcs) modulate RAG-mediated cleavage. Modulation of cleavage by Ku70/80 and DNA-PKcs results in preferential inhibition of 12/12 and 23/23 DNA cleavage, thus increasing 12/23 rule specificity. This observation indicates that DNA repair factors, Ku70/80 and DNA-PKcs, might be present upstream of the DNA cleavage events and not recruited downstream as is currently thought, assigning new nonrepair functions to the DNA-dependent protein kinase.",
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AU - Alarcon, Claudio

AU - Singha, Netai C.

AU - Langen, Hanno

AU - Bianchi, Marco E.

AU - Lees-Miller, Susan P.

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