A microarray-based genetic screen for yeast chronological aging factors.
A microarray-based genetic screen for yeast chronological aging factors.
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DOI：
10.1371/journal.pgen.1000921
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发表时间：
2010-04-22
期刊：
影响因子：
4.5
通讯作者：
Smith JS
中科院分区：
文献类型：
--
作者：
Matecic M;Smith DL;Pan X;Maqani N;Bekiranov S;Boeke JD;Smith JS
模式生物在阐明调节衰老的多个基因和细胞过程中发挥了重要作用。在这项研究中，我们利用芽殖酵母，酿酒酵母，在一个大规模的屏幕上的基因，在调节的时间寿命，这是由非分裂细胞保持活力的天数定义的功能。在液体培养物中按时间顺序老化活的单倍体基因缺失突变体的汇集物，每个突变体用独特的识别DNA“条形码”序列标记。在几个时间点选择老化群体中的活突变体，然后使用定量条形码标签丰度的微阵列DNA杂交技术检测。使用这种方法鉴定了多个短寿命和长寿命突变体。在已证实的短寿命突变体中，有自噬缺陷的突变体，这表明细胞器在长寿中的回收利用是一个关键要求。自噬的缺陷也阻止了由生长培养基中氨基酸的限制诱导的寿命延长。在已证实的长寿突变体中，有高度保守的从头嘌呤生物合成途径（ADE基因）缺陷的突变体，该途径最终产生IMP和AMP。阻断这一途径延长寿命的程度与限制卡路里（葡萄糖）相同。最近发现的一种与醋酸分泌和毒性有关的时间老化的细胞外在机制在一种长寿的ade4Δ突变体中受到抑制，并被一种短命的atg16Δ自噬突变体加剧。酵母按时间顺序的寿命的多个新的效应物的鉴定将大大有助于阐明细胞和生物体在减缓衰老过程中所利用的机制。衰老过程与包括糖尿病和癌症在内的几种与年龄相关的疾病的发病有关。在啮齿动物模型系统中，已知称为热量限制（CR）的饮食方案可以延迟或预防这些疾病并延长寿命。因此，有很大的兴趣，了解CR功能的机制。芽殖酵母，酿酒酵母，已被证明是一个有效的模型，用于分析基因和细胞通路，有助于调节衰老。在这项研究中，我们已经进行了基于微阵列的遗传筛选酵母中，确定短寿命和长寿命的突变体，从一个人口，其中包含每个可行的单倍体基因缺失突变体从酵母基因敲除收集，汇集在一起。使用这种方法，我们能够从以前没有参与衰老的几种途径中鉴定出基因，包括一些似乎有助于限制氨基酸或糖诱导的CR效应的基因。这些结果有望为未来更复杂生物体的机械老化研究提供新的基础。
Model organisms have played an important role in the elucidation of multiple genes and cellular processes that regulate aging. In this study we utilized the budding yeast, Saccharomyces cerevisiae, in a large-scale screen for genes that function in the regulation of chronological lifespan, which is defined by the number of days that non-dividing cells remain viable. A pooled collection of viable haploid gene deletion mutants, each tagged with unique identifying DNA “bar-code” sequences was chronologically aged in liquid culture. Viable mutants in the aging population were selected at several time points and then detected using a microarray DNA hybridization technique that quantifies abundance of the barcode tags. Multiple short- and long-lived mutants were identified using this approach. Among the confirmed short-lived mutants were those defective for autophagy, indicating a key requirement for the recycling of cellular organelles in longevity. Defects in autophagy also prevented lifespan extension induced by limitation of amino acids in the growth media. Among the confirmed long-lived mutants were those defective in the highly conserved de novo purine biosynthesis pathway (the ADE genes), which ultimately produces IMP and AMP. Blocking this pathway extended lifespan to the same degree as calorie (glucose) restriction. A recently discovered cell-extrinsic mechanism of chronological aging involving acetic acid secretion and toxicity was suppressed in a long-lived ade4Δ mutant and exacerbated by a short-lived atg16Δ autophagy mutant. The identification of multiple novel effectors of yeast chronological lifespan will greatly aid in the elucidation of mechanisms that cells and organisms utilize in slowing down the aging process. The aging process is associated with the onset of several age-associated diseases including diabetes and cancer. In rodent model systems, the dietary regimen known as caloric restriction (CR) is known to delay or prevent these diseases and to extend lifespan. As a result, there is a great deal of interest in understanding the mechanisms by which CR functions. The budding yeast, Saccharomyces cerevisiae, has proven to be an effective model for the analysis of genes and cellular pathways that contribute to the regulation of aging. In this study we have performed a microarray-based genetic screen in yeast that identified short- and long-lived mutants from a population that contained each of the viable haploid gene deletion mutants from the yeast gene knockout collection that were pooled together. Using such an approach, we were able to identify genes from several pathways that had not been previously implicated in aging, including some that appear to contribute to the CR effect induced by restriction of either amino acids or sugar. These results are expected to provide new groundwork for future mechanistic aging studies in more complex organisms.
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影响因子：
7.8
作者：
Alvers AL;Fishwick LK;Wood MS;Hu D;Chung HS;Dunn WA Jr;Aris JP
通讯作者：
Aris JP
影响因子：
10.5
作者：
Hamilton, B;Doug, YQ;Lee, SS
通讯作者：
Lee, SS
DOI：
10.1083/jcb.200404002
发表时间：
2004-09-27
期刊：
The Journal of cell biology
影响因子：
--
作者：
Fabrizio P;Battistella L;Vardavas R;Gattazzo C;Liou LL;Diaspro A;Dossen JW;Gralla EB;Longo VD
通讯作者：
Longo VD
影响因子：
56.9
作者：
Fabrizio, P;Pozza, F;Longo, VD
通讯作者：
Longo, VD
影响因子：
4.5
作者：
Hansen M;Hsu AL;Dillin A;Kenyon C
通讯作者：
Kenyon C