Signal joint formation is also impaired in DNA-dependent protein kinase catalytic subunit knockout cells

R. Fukumura, R. Araki, A. Fujimori, Y. Tsutsumi, A. Kurimasa, G. C. Li, D. J. Chen, K. Tatsumi, M. Abe

Research output: Contribution to journalArticle

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Abstract

The effort to elucidate the mechanism of V(D)J recombination has given rise to a dispute as to whether DNA-dependent protein kinase catalytic subunit (DNA-PKcs) contributes to signal joint formation (sjf). Observations reported to date are confusing. Analyses using DNA-PKcs-deficient cells could not conclude the requirement of DNA-PKcs for sjf, because sjf can be formed by end-joining activities which are diverse among cells other than those participating in V(D)J recombination. Here, we observed V(D)J recombination in DNA-PKcs knockout cells and showed that both signal and coding joint formation were clearly impaired in the cells. Subsequently, to directly demonstrate the requirement of DNA-PKcs for sjf, we introduced full-length cDNA of DNA-PKcs into the knockout cells. Furthermore, several mutant DNA-PKcs cDNA constructs designed from mutant cell lines (irs-20, V3, murine scid, and SX9) were also introduced into the cells to obtain further evidence indicating the involvement of DNA-PKcs in sjf. We found as a result that the full-length cDNA complemented the aberrant sjf and that the mutant cDNAs constructs also partially complemented it. Lastly, we looked at whether the kinase activity of DNA-PKcs is necessary for sjf and, as a result, demonstrated a close relationship between them. Our observations clearly indicate that the DNA-PKcs controls not only coding joint formation but also the sjf in V(D)J recombination through its kinase activity.

Original languageEnglish (US)
Pages (from-to)3883-3889
Number of pages7
JournalJournal of Immunology
Volume165
Issue number7
StatePublished - Oct 1 2000

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DNA-Activated Protein Kinase
Catalytic Domain
Joints
V(D)J Recombination
Complementary DNA
Phosphotransferases
Dissent and Disputes

ASJC Scopus subject areas

  • Immunology

Cite this

Fukumura, R., Araki, R., Fujimori, A., Tsutsumi, Y., Kurimasa, A., Li, G. C., ... Abe, M. (2000). Signal joint formation is also impaired in DNA-dependent protein kinase catalytic subunit knockout cells. Journal of Immunology, 165(7), 3883-3889.

Signal joint formation is also impaired in DNA-dependent protein kinase catalytic subunit knockout cells. / Fukumura, R.; Araki, R.; Fujimori, A.; Tsutsumi, Y.; Kurimasa, A.; Li, G. C.; Chen, D. J.; Tatsumi, K.; Abe, M.

In: Journal of Immunology, Vol. 165, No. 7, 01.10.2000, p. 3883-3889.

Research output: Contribution to journalArticle

Fukumura, R, Araki, R, Fujimori, A, Tsutsumi, Y, Kurimasa, A, Li, GC, Chen, DJ, Tatsumi, K & Abe, M 2000, 'Signal joint formation is also impaired in DNA-dependent protein kinase catalytic subunit knockout cells', Journal of Immunology, vol. 165, no. 7, pp. 3883-3889.
Fukumura R, Araki R, Fujimori A, Tsutsumi Y, Kurimasa A, Li GC et al. Signal joint formation is also impaired in DNA-dependent protein kinase catalytic subunit knockout cells. Journal of Immunology. 2000 Oct 1;165(7):3883-3889.
Fukumura, R. ; Araki, R. ; Fujimori, A. ; Tsutsumi, Y. ; Kurimasa, A. ; Li, G. C. ; Chen, D. J. ; Tatsumi, K. ; Abe, M. / Signal joint formation is also impaired in DNA-dependent protein kinase catalytic subunit knockout cells. In: Journal of Immunology. 2000 ; Vol. 165, No. 7. pp. 3883-3889.
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