| Co-Investigator(Kenkyū-buntansha) |
KANAI Fumihiko The University of Tokyo, Faculty of Medicine, visiting Assistant Professor, 医学部附属病院, 客員助教授 (70334399)
OGURA Keiji The University of Tokyo, Faculty of Medicine, Assistant Professor, 医学部附属病院, 助手 (50376456)
|
|---|
| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2006: ¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 2005: ¥1,800,000 (Direct Cost: ¥1,800,000)
|
|---|
| Research Abstract |
Recent studies have suggested that Runt-related transcription factor 3(Runx3) is associated with genesis and progression of gastric carcinoma. To elucidate the molecular mechanisms of gastric carcinogenesis, a proteomic approach was used to search for Runx3-interacting proteins. Runx3 bound with myc and flag tags is expressed in HEK293T cells, and the protein complex formed with Runx3 was purified and identified by mass spectrometry. Ku70 and Ku80, members of the DNA repair protein complex, were identified as Runx3-interacting proteins. Runx3, Ku70, and Ku80 associate in vivo, and direct interaction between Runx3 and Ku70 was confirmed via His-tag pull-down assay. The amino acids 241-322 of Runx3, which correspond to the transcriptional activation domain, and the amino acids 1-116 of Ku70 were necessary for binding with each other, and immunocytochemistry under confocal laser microscopy demonstrated that Runx3 and Ku70 colocalized throughout the nucleus.
We also analyzed the effect of Ku70 to Runx3 transcriptional regulation. We performed luciferase assay using p21 and Runx reporters in Ku70 knockdown cells. However knockdown of Ku70 did not affect p21 or Runx regulation significantly.
Moreover, we investigate the association between Runx3 and TAZ, that is a novel transcriptional activator, and examine the expression level of Runx3 by using clinical samples(chronic gastritis, atrophic gastritis and gastric cancer), though the number of samples analyzed was too small to conclude the association between Runx3 and gastric disease, thus further investigation is needed.
|
