Drosophila protein kinase CK2 is rendered temperature-sensitive by mutations of highly conserved residues flanking the activation segment.

Drosophila protein kinase CK2 is rendered temperature-sensitive by mutations of highly conserved residues flanking the activation segment.
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DOI:
10.1007/s11010-008-9963-6
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发表时间:
2009-03
影响因子:
4.3
通讯作者:
Bidwai, Ashok P.
Bidwai, Ashok P.
中科院分区:
生物学3区
文献类型:
--
作者:
Kuntamalla, Pallavi P.;Kunttas-Tatli, Ezgi;Karandikar, Umesh;Bishop, Clifton P.;Bidwai, Ashok P.

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CK2是动物发育所必需的丝氨酸/苏氨酸蛋白激酶。尽管在小鼠和果蝇模型中存在CK2的零等位基因,但它们在纯合时是致命的,因此需要条件等位基因来分析其发育作用。我们描述了果蝇CK2α (dCK2α)温度敏感等位基因的分离。这些等位基因在29°C时有效地挽救了缺乏内源性CK2的酵母的致死率,但这种能力在更高的温度下以等位基因特异性的方式丧失。这些ts-变体表现出与野生型蛋白相似的特性,并与dCK2β强烈相互作用。利用人类CK2α的晶体结构对这些ts-变体进行建模表明,受影响的残基非常接近活性位点。我们发现Asp212的替代引发了强有力的ts行为,这是一个重要的发现,因为该残基有助于激活片段的稳定性,并且在其他丝氨酸/苏氨酸蛋白激酶中是不变的。
CK2 is a Ser/Thr protein kinase essential for animal development. Although null alleles for CK2 are available in the mouse and Drosophila models, they are lethal when homozygous, thus necessitating conditional alleles for analysis of its developmental roles. We describe the isolation of temperature-sensitive (ts) alleles of Drosophila CK2α (dCK2α). These alleles efficiently rescue lethality of yeast lacking endogenous CK2 at 29°C, but this ability is lost at higher temperatures in an allele-specific manner. These ts-variants exhibit properties akin to the wild type protein, and interact robustly with dCK2β. Modeling of these ts-variants using the crystal structure of human CK2α indicates that the affected residues are in close proximity to the active site. We find that substitution of Asp212 elicits potent ts-behavior, an important finding because this residue contributes to stability of the activation segment and is invariant in other Ser/Thr protein kinases.
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