DDB1 gene disruption causes a severe growth defect and apoptosis in chicken DT40 cells

Mitsuo Wakasugi, Kenkyo Matsuura, Atsushi Nagasawa, DongTao Fu, Hiroko Shimizu, Ken-ichi Yamamoto, Shunichi Takeda, Tsukasa Matsunaga

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

DDB1 was originally identified as a heterodimeric complex with DDB2 and plays an accessory role in nucleotide excision repair. DDB1 also constitutes an E3 ubiquitin ligase complex together with Cul4A and Roc1 and acts as an adaptor, suggesting its multiple roles beyond DNA repair. We have generated a conditional DDB1-knockout mutant using a chicken B lymphocyte line DT40. Doxycycline-induced DDB1 depletion caused a severe growth defect followed by apoptotic cell death. Flow cytometric analyses revealed that cell cycle progression is initially retarded at all phases and subsequently impaired at S phase along with the appearance of sub-G1 population. Similarly, DDB1-knockdown in human U2OS cells by small interfering RNA exhibited a loss of clonogenic activity and perturbed cell cycle progression. These results demonstrate that the DDB1 gene is indispensable for cell viability in higher vertebrates and this conditional DDB1-knockout clone would be highly useful for the functional analysis of DDB1.

Original languageEnglish
Pages (from-to)771-777
Number of pages7
JournalBiochemical and Biophysical Research Communications
Volume364
Issue number4
DOIs
Publication statusPublished - Dec 28 2007
Externally publishedYes

Fingerprint

DNA Repair
Chickens
Cell Cycle
Genes
Cells
Apoptosis
Defects
Ubiquitin-Protein Ligases
Doxycycline
Repair
Growth
S Phase
Small Interfering RNA
Vertebrates
Cell Survival
Functional analysis
B-Lymphocytes
Cell Death
Lymphocytes
Clone Cells

Keywords

  • Apoptosis
  • Cell cycle arrest
  • Cell viability
  • Conditional knockout
  • DDB1
  • DT40

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

DDB1 gene disruption causes a severe growth defect and apoptosis in chicken DT40 cells. / Wakasugi, Mitsuo; Matsuura, Kenkyo; Nagasawa, Atsushi; Fu, DongTao; Shimizu, Hiroko; Yamamoto, Ken-ichi; Takeda, Shunichi; Matsunaga, Tsukasa.

In: Biochemical and Biophysical Research Communications, Vol. 364, No. 4, 28.12.2007, p. 771-777.

Research output: Contribution to journalArticle

Wakasugi, Mitsuo ; Matsuura, Kenkyo ; Nagasawa, Atsushi ; Fu, DongTao ; Shimizu, Hiroko ; Yamamoto, Ken-ichi ; Takeda, Shunichi ; Matsunaga, Tsukasa. / DDB1 gene disruption causes a severe growth defect and apoptosis in chicken DT40 cells. In: Biochemical and Biophysical Research Communications. 2007 ; Vol. 364, No. 4. pp. 771-777.
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