Telomere-binding and Stn1p-interacting activities are required for the essential function of Saccharomyces cerevisiae Cdc13p.
Telomere-binding and Stn1p-interacting activities are required for the essential function of Saccharomyces cerevisiae Cdc13p.
复制标题
端粒结合和 Stn1p 相互作用活性是酿酒酵母 Cdc13p 基本功能所必需的。
作者：
Mei;Yi;Te;Jui;Chia;Jing
酿酒酵母Cdc 13 p是一种端粒结合蛋白，它能保护端粒并调节端粒长度。据记载，Cdc 1 - 3 p特异性结合到单链TG（1-3）端粒DNA序列，并与Stn 1 p相互作用。为了定位单链TG（1-3）DNA结合区域，通过缺失突变构建了Cdc 13 p突变体并测定了其结合活性。基于体外电泳迁移率变动分析，Cdc 1 - 3 p的243个氨基酸片段（氨基酸451-693）足以结合单链TG（1-3），其特异性与天然蛋白相似。与体外观察一致，体内单杂交分析也表明Cdc 13 p的这一区域足以将其自身定位于端粒。然而，Cdc 13 p的端粒结合区（氨基酸451-693）不能补充cdc 13突变体的生长缺陷。相反，包含Cdc 13 p的Stn 1 p相互作用和端粒结合区（氨基酸252-924）的区域补充了cdc 13突变体的生长缺陷。这些结果表明Cdc 13 p与端粒的结合不足以解释细胞活力，还需要与Stn 1 p的相互作用。两者合计，我们已经定义了Cdc 13 p的端粒结合结构域，并表明Cdc 13 p与端粒和Stn 1 p的结合是维持细胞生长所必需的。
Yeast Saccharomyces cerevisiae Cdc13p is the telomere-binding protein that protects telomeres and regulates telomere length. It is documented that Cdc13p binds specifically to single-stranded TG(1-3) telomeric DNA sequences and interacts with Stn1p. To localize the region for single-stranded TG(1-3) DNA binding, Cdc13p mutants were constructed by deletion mutagenesis and assayed for their binding activity. Based on in vitro electrophoretic mobility shift assay, a 243-amino-acid fragment of Cdc13p (amino acids 451-693) was sufficient to bind single-stranded TG(1-3) with specificity similar to that of the native protein. Consistent with the in vitro observation, in vivo one-hybrid analysis also indicated that this region of Cdc13p was sufficient to localize itself to telomeres. However, the telomere-binding region of Cdc13p (amino acids 451-693) was not capable of complementing the growth defects of cdc13 mutants. Instead, a region comprising the Stn1p-interacting and telomere-binding region of Cdc13p (amino acids 252-924) complemented the growth defects of cdc13 mutants. These results suggest that binding to telomeres by Cdc13p is not sufficient to account for the cell viability, interaction with Stn1p is also required. Taken together, we have defined the telomere-binding domain of Cdc13p and showed that both binding to telomeres and Stn1p by Cdc13p are required to maintain cell growth.
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影响因子：
10.5
作者：
VirtaPearlman, V;Morris, DK;Lundblad, V
通讯作者：
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影响因子：
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影响因子：
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作者：
Stavenhagen,JB;Zakian,VA
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影响因子：
5.6
作者：
Prasad,BV;Chiu,W
通讯作者：
Chiu,W
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14.9
作者：
J. Wright;V. Zakian
通讯作者：
J. Wright;V. Zakian