2000 Fiscal Year Final Research Report Summary
Regulatory mechanism of cellular functions of yeast by the ubiquitin-proeasome system.
| Project/Area Number |
10440225
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (B).
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
遺伝
|
|---|
| Research Institution | The University of Tokyo |
|---|
Principal Investigator |
TOH-E Akio The University of Tokyo, Department of Biological Sciences, Professor, 大学院・理学系研究科, 教授 (90029249)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
UESONO Yukifumi The University of Tokyo, Department of Biological Sciences, Assistant Professor, 大学院・理学系研究科, 助手 (30251408)
KIKUCHI Yoshiko The University of Tokyo, Department of Biological Sciences, Associated, 大学院・理学系研究科, 助教授 (00138124)
|
|---|
| Project Period (FY) |
1998 – 2000
|
| Keywords | 26S proteasome / cell cycle / mitosis / metaphase-anaphase transition / RPN9 / Saccharomyces cerevisiae / two-hybrid screening |
|---|
| Research Abstract |
We have isolated the RPN9 gene by two hybrid screening using RPN10 (formerly SUN1) as bait, which encodes a multiubiquitin chain receptor residing in the 26S proteasome. A yeast strain carrying a disrupted allele of RPN9 was found to be temperature sensitive for its growth. At the restrictive temperature, the Δrpn9 strain accumulated multiubiquitinated proteins. When the proteasome fractions separated by the glycerol gradient centrifugation were analysed by native PAGE, we found that the 26S proteasome of the Δrpn9 cells was shifted to lighter fractions than expected and that there was a larger amount of the incomplete proteasome complexes in the fractions. Furthermore, Rpn10 was not detected in the 26S proteasome of Δrpn9 cells. These results indicate that Rpn9 is needed for incorporating Rpn10 into the 26S proteasome and that Rpn9 participates in assembly and/or stability of the 26S proteasome.
Δrpn9 cells were found to be arrested at metaphase ; dumbbell shaped, single nucleus at the isthmuth, unseparated sister chromatids. When a Δrpn9 Δpds1 strain was shifted to a restrictive temperature, the cells stopped growing. No anaphase cells were observed, but sister chrmomatids were separated. Judging from these phenotypes, we anticipate the presence of some proteins which should be degraded by the 26S proteasome during anaphase progression.
|
-
-
-
-
-
-
-
-
[Publications] Hori, T., Kato, S., Saeki, M., DeMartino, G.N., Slaughter, C.A., Takeuchi, J., Toh-e, A.& Tanaka, K.: "cDNA cloning and functional analysis of p28 (Nas6p) and p40.5 (Nas7p), two novel regulatory subunits of the 26S proteasome."Gene. 216. 113-122 (1998)
Description
「研究成果報告書概要(欧文)」より
-
-
[Publications] Fujimuro, M., Takada, H., Saeki, Y., Toh-e, A., Tanaka, K.& Yokosawa, H.: "Growth-dependent change of the 26S proteasome in budding yeast."Biochem.Biophys.Res.Commun.. 251. 818-823 (1998)
Description
「研究成果報告書概要(欧文)」より
-
-
[Publications] Takahashi, Y., Iwase, M., Konishi, M., Tanaka, M., Toh-e, A.& Kikuchi, Y.: "Smt3, a SUMO-1 homolog, is conjugated to Cdc3, a component of septin rings at the mother-budneck in budding yeast."Biochem.Biophys.Res.Commun.. 259. 582-587 (1999)
Description
「研究成果報告書概要(欧文)」より
-
-
-
[Publications] Tone, Y., Tanahashi, N., Tanaka, K., Fujimura, M., Yokosawa, H.& Toh-e, A.: "Nob1p, a new essential protein, associates with the 26S proteasome of growing Saccharomyces cerevisiae cells."Gene. 243. 37-45 (2000)
Description
「研究成果報告書概要(欧文)」より
-
-
-
-
[Publications] Nishizawa, M., Tanabe, M., Yabuki, N., Kitada, K., & Toh-e, A.: "Pho85 kinase, a yeast cyclin-dependent kinase, regulates the expession of UGP1 encoding UDP-glucose pyrophosphorylase."Yeast. (in press). (2001)
Description
「研究成果報告書概要(欧文)」より
-
