| Project/Area Number |
06044197
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| Research Category |
Grant-in-Aid for international Scientific Research
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| Allocation Type | Single-year Grants |
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| Section | Joint Research |
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| Research Institution | Keio University School of Medicine |
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Principal Investigator |
FUKASAWA Toshio Keio University, School of Medicine, Professor, 医学部, 名誉教授 (90029934)
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| Co-Investigator(Kenkyū-buntansha) |
SUZUKI Yuriko Keio University School of Medicine, 医学部, 助手 (40255435)
LORCH Yahli Stanford University school of Medicine, 医学部, 博士研究員
JOHNSTON Stephen a University of Texas Southwetern Medical Center, 医学部, 準教授
KORNBERG Roger d Stanford University School of Medicine, 医学部, 教授
WADA Tadashi Tokyo Institute of Technology, 生命理工学部, 助手 (60262309)
HANDA Hiroshi Tokyo Institute of Technology, 生命理工学部, 教授 (80107432)
SAKURAI Hiroshi Keio University School of Medicine, 医学部, 助手 (00225848)
BERK A.J. カリフォルニア大学ロスアンゼルス校, 分子生物研究所, 教授
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| Project Period (FY) |
1994 – 1995
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| Project Status |
Completed (Fiscal Year 1995)
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| Budget Amount *help |
¥6,100,000 (Direct Cost: ¥6,100,000)
Fiscal Year 1995: ¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 1994: ¥3,100,000 (Direct Cost: ¥3,100,000)
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| Keywords | Galactose / Reconstitution System / Basal transcription / Basal factor / Phosphoryration / Saccharomyces cerevisiae |
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| Research Abstract |
1. Analysis of galactose signal transduction
The GAL3 gene plays an critical role in galactose induction of the genes encoding galactose-metabolizing enzymes (GAL) in yeast. Defects in GAL3 result in a long delay in inducing GAL genes, and overproduction of Gal3p causes constitutive expression of GAL.Here we demonstrate concomitant overproduction of the negative regulator, Gal80p, and Gal3p suppresses this GAL constitutive expression. This interplay between Gal80p and Gal3p is direct as tagged Gal3p co-immunoprecipitates with Gal80p. The amount of coprecipitated Gal80p increases when GAL80 yeast is grown in the presence of galactose. When both GAL80 and GAL3 are overexpressed, the amount of coprecipitated Gal80p is not affected by galactose. Tagged gal3 mutant proteins bind to purified Gal80p but only poorly in comparison to the wild type, suggesting that the formation of the Gal80p/Gal3p complex depends on the normal function of Gal3p. The Gal3p appeared larger in western blots than pr
… More
edicted by the published nucleic acid sequence.
2. Interaction between specific transcription factors and basal transcription machinery
The GAL11 gene encodes an auxiliary transcription factor required for full expression of most yeast genes except those whose expression is independent of the canonical TATA box so far as studied. The GAL11 encoded protein (Gal11p) has recently been shown to copurify with the holoenzyme of RNA polymerase II.In fact, immunoprecipitation experiments with anti-Gal11p antibody in our laboratory show that Gal11p coprecipitates with the largest subunit of RNA polymerase II.Here we report that Gal11p stimulates the basal transcription in a reconstituted transcription system composed of recombinant or highly purified transcription factors (TF), IIB,IIE,IIF,IIH,and TBP and core RNA polymerase II.We further demonstrate that Gal11p directly binds TFIIE.Each of the two domains of Gal11p essential for in vivo function respectively participates in the binding to the small and large subunits of TFIIE.In light of these findings, we suggest that Gal11p modulates the promoter activity of the TATA-bearing genes by recruiting TFIIE to the preinitiation complex in yeast. Less
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