Investigating the role of the Est3 protein in yeast telomere replication.
Investigating the role of the Est3 protein in yeast telomere replication.
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DOI：
10.1093/nar/gkp1173
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发表时间：
2010-04
影响因子：
14.9
通讯作者：
Lundblad V
中科院分区：
文献类型：
--
作者：
Lee J;Mandell EK;Rao T;Wuttke DS;Lundblad V
酵母端粒酶的Est 3亚基采用预测的OB折叠，对于端粒复制至关重要。为了评估Est 3可能对酶催化的贡献，我们比较了来自野生型和Castellii酵母的est 3-Δ菌株的端粒酶活性，结果显示Est 3亚基的缺失导致核苷酸添加减少2至3倍。基于对跨越S.卡氏端粒酶RNA。此外，使用核磁共振化学位移扰动，没有观察到化学位移的变化，在任何网站上的蛋白质后，添加单链DNA，反对的作用Est 3在端粒底物的端粒酶识别。添加外源性Est 3蛋白，包括在体内端粒复制严重受损的突变Est 3蛋白，完全恢复了est 3-Δ端粒酶反应的活性。因此，Est 3在端粒复制中执行体内调节功能，这与Est 3可能对端粒酶活性做出的任何潜在贡献不同。
The Est3 subunit of yeast telomerase, which adopts a predicted OB-fold, is essential for telomere replication. To assess the possible contributions that Est3 might make to enzyme catalysis, we compared telomerase activity from wild type and est3-Δ strains of Saccharomyces castellii, which revealed that loss of the Est3 subunit results in a 2- to 3-fold decline in nucleotide addition. This effect was not primer-specific, based on assessment of a panel of primers that spanned the template of the S. castellii telomerase RNA. Furthermore, using nuclear magnetic resonance chemical shift perturbation, no chemical shift change was observed at any site in the protein upon addition of single-stranded DNA, arguing against a role for Est3 in recognition of telomeric substrates by telomerase. Addition of exogenous Est3 protein, including mutant Est3 proteins that are severely impaired for telomere replication in vivo, fully restored activity in est3-Δ telomerase reactions. Thus, Est3 performs an in vivo regulatory function in telomere replication, which is distinct from any potential contribution that Est3 might make to telomerase activity.
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DOI：
10.1073/pnas.0700327104
发表时间：
2007-07-10
影响因子：
11.1
作者：
Hsu, Min;McEachern, Michael J.;Lue, Neal F.
通讯作者：
Lue, Neal F.
影响因子：
5.3
作者：
LEE, MS;BLACKBURN, EH
通讯作者：
BLACKBURN, EH
影响因子：
56.9
作者：
COHN, M;BLACKBURN, EH
通讯作者：
BLACKBURN, EH
影响因子：
11.4
作者：
Marcand, S;Brevet, V;Gilson, E
通讯作者：
Gilson, E
影响因子：
9.2
作者：
Dandjinou, AT;Lévesque, N;Wellinger, RJ
通讯作者：
Wellinger, RJ