Perturbation of the yeast N-acetyltransferase NatB induces elevation of protein phosphorylation levels.
Perturbation of the yeast N-acetyltransferase NatB induces elevation of protein phosphorylation levels.
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DOI：
10.1186/1471-2164-11-685
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发表时间：
2010-12-02
期刊：
影响因子：
4.4
通讯作者：
Heck AJ
中科院分区：
文献类型：
--
作者：
Helbig AO;Rosati S;Pijnappel PW;van Breukelen B;Timmers MH;Mohammed S;Slijper M;Heck AJ
在蛋白质N-末端添加乙酰基是一种广泛的共翻译修饰。NatB是主要的N-乙酰转移酶之一，其靶向具有N-末端甲硫氨酸的蛋白质的子集，但到目前为止仅报道了少数底物。使用酵母nat 3 Δ菌株，缺乏NatB的催化亚基，我们采用定量蛋白质组学策略来鉴定NatB底物并表征nat 3 Δ中的下游效应。通过蛋白质组学比较WT和nat 3 Δ菌株，使用代谢15 N同位素标记，我们确信地在总共756个检测到的乙酰化蛋白N-末端中鉴定了59个NatB底物。我们获得了大约2580种蛋白质表达水平的深入蛋白质组测量。最显著的是，NatB缺失导致蛋白质磷酸化的非常显著的变化。蛋白质表达水平在WT和nat 3 Δ之间仅略微变化。检测到的NatB基板与他们的orthophosphate的比较显示在整个系统发育树的保护非常少。我们进一步提出的证据，翻译后N-乙酰化的蛋白质变体在非注释的N-末端。此外，对nat 3 Δ下游效应的分析显示，蛋白磷酸化水平升高，因此激酶Snf 1 p可能是该过程中的关键因素。
The addition of an acetyl group to protein N-termini is a widespread co-translational modification. NatB is one of the main N-acetyltransferases that targets a subset of proteins possessing an N-terminal methionine, but so far only a handful of substrates have been reported. Using a yeast nat3Δ strain, deficient for the catalytic subunit of NatB, we employed a quantitative proteomics strategy to identify NatB substrates and to characterize downstream effects in nat3Δ. Comparing by proteomics WT and nat3Δ strains, using metabolic 15N isotope labeling, we confidently identified 59 NatB substrates, out of a total of 756 detected acetylated protein N-termini. We acquired in-depth proteome wide measurements of expression levels of about 2580 proteins. Most remarkably, NatB deletion led to a very significant change in protein phosphorylation. Protein expression levels change only marginally in between WT and nat3Δ. A comparison of the detected NatB substrates with their orthologous revealed remarkably little conservation throughout the phylogenetic tree. We further present evidence of post-translational N-acetylation on protein variants at non-annotated N-termini. Moreover, analysis of downstream effects in nat3Δ revealed elevated protein phosphorylation levels whereby the kinase Snf1p is likely a key element in this process.
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影响因子：
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