Loss of APD1 in yeast confers hydroxyurea sensitivity suppressed by Yap1p transcription factor.
Loss of APD1 in yeast confers hydroxyurea sensitivity suppressed by Yap1p transcription factor.
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酵母中 APD1 的缺失导致羟基脲敏感性受到 Yap1p 转录因子的抑制。
DOI：
10.1038/srep07897
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发表时间：
2015-01-20
影响因子：
4.6
通讯作者：
Jin DY
中科院分区：
文献类型：
--
作者：
Tang HM;Pan K;Kong KY;Hu L;Chan LC;Siu KL;Sun H;Wong CM;Jin DY
铁还原蛋白是在电子传递和氧化还原稳态中起重要作用的铁硫蛋白。酵母Apd1p是类硫氧还蛋白家族的新成员。在这项研究中，我们表征了apd1Δ细胞的羟基脲（HU）超敏表型。胡是一种DNA合成抑制剂、细胞应激源和抗癌剂。虽然APD1的缺失不影响细胞增殖或细胞周期进展，但会导致HU敏感性。这种敏感性在抗氧化剂n -乙酰半胱氨酸的存在下恢复，暗示了细胞内氧化还原的作用。Apd1p中铁结合基序的突变使其在apd1Δ细胞中恢复HU敏感性的能力丧失。Apd1p的铁结合活性通过显色法验证。通过质谱分析，发现两个铁被合并到一个Apd1p蛋白分子中。令人惊讶的是，核糖核苷酸还原酶基因在apd1Δ细胞中没有被诱导，当dNTP产量增加时，HU敏感性不受影响。通过抑制因子筛选，发现应激调节转录因子Yap1p的表达可以有效地挽救apd1Δ细胞的HU敏感性。综上所述，我们的工作确定了Apd1p是一种新的铁氧还蛋白，在细胞防御HU中起关键作用。
Ferredoxins are iron-sulfur proteins that play important roles in electron transport and redox homeostasis. Yeast Apd1p is a novel member of the family of thioredoxin-like ferredoxins. In this study, we characterized the hydroxyurea (HU)-hypersensitive phenotype of apd1Δ cells. HU is an inhibitor of DNA synthesis, a cellular stressor and an anticancer agent. Although the loss of APD1 did not influence cell proliferation or cell cycle progression, it resulted in HU sensitivity. This sensitivity was reverted in the presence of antioxidant N-acetyl-cysteine, implicating a role for intracellular redox. Mutation of the iron-binding motifs in Apd1p abrogated its ability to rescue HU sensitivity in apd1Δ cells. The iron-binding activity of Apd1p was verified by a color assay. By mass spectrometry two irons were found to be incorporated into one Apd1p protein molecule. Surprisingly, ribonucleotide reductase genes were not induced in apd1Δ cells and the HU sensitivity was unaffected when dNTP production was boosted. A suppressor screen was performed and the expression of stress-regulated transcription factor Yap1p was found to effectively rescue the HU sensitivity in apd1Δ cells. Taken together, our work identified Apd1p as a new ferredoxin which serves critical roles in cellular defense against HU.
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影响因子：
3.1
作者：
Dubacq, C;Chevalier, A;Mann, C
通讯作者：
Mann, C
DOI：
10.1007/pl00013817
发表时间：
1999-12-01
期刊：
MOLECULAR AND GENERAL GENETICS
影响因子：
--
作者：
Entian, KD;Schuster, T;Hinnen, A
通讯作者：
Hinnen, A
DOI：
10.1073/pnas.0610585104
发表时间：
2007-01-23
影响因子：
11.1
作者：
Chabes, Andrei;Stillman, Bruce
通讯作者：
Stillman, Bruce
影响因子：
64.5
作者：
Chabes, A;Georgieva, B;Thelander, L
通讯作者：
Thelander, L
影响因子：
64.5
作者：
Huang, MX;Zhou, Z;Elledge, SJ
通讯作者：
Elledge, SJ