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一项针对酿酒酵母中参与感知无机磷酸盐可用性的基因的系统性遗传筛选。

A systematic genetic screen for genes involved in sensing inorganic phosphate availability in Saccharomyces cerevisiae.

作者信息

Choi Joonhyuk, Rajagopal Abbhirami, Xu Yi-Fan, Rabinowitz Joshua D, O'Shea Erin K

机构信息

Faculty of Arts and Sciences Center for Systems Biology, Harvard University, Cambridge, Massachusetts, United States of America.

Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts, United States of America.

出版信息

PLoS One. 2017 May 17;12(5):e0176085. doi: 10.1371/journal.pone.0176085. eCollection 2017.

DOI:10.1371/journal.pone.0176085
PMID:28520786
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5435139/
Abstract

Saccharomyces cerevisiae responds to changes in extracellular inorganic phosphate (Pi) availability by regulating the activity of the phosphate-responsive (PHO) signaling pathway, enabling cells to maintain intracellular levels of the essential nutrient Pi. Pi-limitation induces upregulation of inositol heptakisphosphate (IP7) synthesized by the inositol hexakisphosphate kinase Vip1, triggering inhibition of the Pho80/Pho85 cyclin-cyclin dependent kinase (CDK) complex by the CDK inhibitor Pho81, which upregulates the PHO regulon through the CDK target and transcription factor Pho4. To identify genes that are involved in signaling upstream of the Pho80/Pho85/Pho81 complex and how they interact with each other to regulate the PHO pathway, we performed genome-wide screens with the synthetic genetic array method. We identified more than 300 mutants with defects in signaling upstream of the Pho80/Pho85/Pho81 complex, including AAH1, which encodes an adenine deaminase that negatively regulates the PHO pathway in a Vip1-dependent manner. Furthermore, we showed that even in the absence of VIP1, the PHO pathway can be activated under prolonged periods of Pi starvation, suggesting complexity in the mechanisms by which the PHO pathway is regulated.

摘要

酿酒酵母通过调节磷酸盐应答（PHO）信号通路的活性来应对细胞外无机磷酸盐（Pi）可用性的变化，使细胞能够维持必需营养素Pi的细胞内水平。Pi限制会诱导由肌醇六磷酸激酶Vip1合成的肌醇七磷酸（IP7）上调，从而触发CDK抑制剂Pho81对Pho80/Pho85细胞周期蛋白-细胞周期蛋白依赖性激酶（CDK）复合物的抑制，该复合物通过CDK靶点和转录因子Pho4上调PHO调节子。为了鉴定参与Pho80/Pho85/Pho81复合物上游信号传导的基因，以及它们如何相互作用以调节PHO途径，我们使用合成遗传阵列方法进行了全基因组筛选。我们鉴定出300多个在Pho80/Pho85/Pho81复合物上游信号传导存在缺陷的突变体，包括AAH1，它编码一种腺嘌呤脱氨酶，以Vip1依赖性方式负调节PHO途径。此外，我们表明，即使在没有VIP1的情况下，PHO途径在长时间的Pi饥饿下也可以被激活，这表明PHO途径的调节机制很复杂。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/57a2/5435139/ced1adb13a12/pone.0176085.g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/57a2/5435139/47c1c5df2aad/pone.0176085.g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/57a2/5435139/2cda7b280f84/pone.0176085.g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/57a2/5435139/809f9af47a11/pone.0176085.g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/57a2/5435139/a3d552eecefb/pone.0176085.g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/57a2/5435139/ccb14ee56c19/pone.0176085.g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/57a2/5435139/ced1adb13a12/pone.0176085.g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/57a2/5435139/47c1c5df2aad/pone.0176085.g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/57a2/5435139/2cda7b280f84/pone.0176085.g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/57a2/5435139/809f9af47a11/pone.0176085.g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/57a2/5435139/a3d552eecefb/pone.0176085.g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/57a2/5435139/ccb14ee56c19/pone.0176085.g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/57a2/5435139/ced1adb13a12/pone.0176085.g006.jpg

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