Structural Functional Analysis of UBF and Fish UBF Gene Identification

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Project title

Code/計畫編號

NSC84-2611-B019-001

Translated Name/計畫中文名

UBF蛋白結構與功能分析及魚類UBF基因鑑認

Project Coordinator/計畫主持人

Chin-Hwa Hu

Funding Organization/主管機關

National Science and Technology Council

Department/Unit

Department of Bioscience and Biotechnology

Website

https://www.grb.gov.tw/search/planDetail?id=162271

Year

1995

Start date/計畫起

01-08-1994

Expected Completion/計畫迄

01-07-1955

Bugetid/研究經費

322千元

ResearchField/研究領域

生物技術(農)

真核生物細胞第一類核酸轉錄酵素(RNA polymerase I)之啟動子UBF具有特殊蛋白耦合結構。在其N-端100個胺基酸序列中,含有四個雙極性.alpha.-螺旋結構(Amphipathic .alpha.-helix)疏水性與親水性胺基酸分列於螺旋結構相對兩側,而此基因啟動子蛋白即藉由兩分子間多個雙極性.alpha.-螺旋結構上之疏水性胺基酸的彼此作用進行蛋白耦合。將此類疏水性胺基酸以定位突變法進行置換,絕大部分均會破壞蛋白耦合之機制,並影響啟動子與DNA間之作用力。 The upstream binding factor, UBF, is an RNA polymerase 1 transcription factor which contains multiple DNA binding domains and a novel protein dimerization domain. Active UBF forms homodimers in vivo through the intramolecular interactions of its dimerization domain, which spans the a hundred amino-terminal residues. In the presence of both UBF dimerization domain and its immediately adjacent lysine-rich basic DNA binding domain, the E. coli expressed recombinant polypeptide, dbUBF (dimerization plus basic motifs of UBF), forms homodimers in vitro and binds to double-stranded DNA nonselectively. In gel retardation assay, dbUBF dimers make multiple shift-ladders corresponding to numerous protein dimer-DNA complexes. The UBF dimerization domain contains multiple helical structures, as predicated by EMBO-PHD program. Most of hydrophobic residues in the dimerization domain are confined in the hydrophobic phase of these hypothetic helices. Mutating these hydrophobic residues to glutamate prohibits dbUBF association and gives different shift pattern in gel retardation assay. The results we present here argues that UBF association is largely exerted by the hydrophobic interactions between the multiple helices to bring two molecules together.

Keyword(s)

蛋白偶合
基因鑑定
Protein dimerization
Gene identification
Upstream binding factor (UBF)

DSpace CRIS

Structural Functional Analysis of UBF and Fish UBF Gene Identification

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