A systematic genetic screen for genes involved in sensing inorganic phosphate availability in Saccharomyces cerevisiae
Abstract
Saccharomyces cerevisiae responds to changes in extracellular inorganic phosphate (P_i) availability by regulating the activity of the phosphate-responsive (PHO) signaling pathway, enabling cells to maintain intracellular levels of the essential nutrient P_i. P_i-limitation induces upregulation of inositol heptakisphosphate (IP_7) 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 P_i starvation, suggesting complexity in the mechanisms by which the PHO pathway is regulated.
Additional Information
© 2017 Choi et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Received: February 13, 2017; Accepted: April 5, 2017; Published: May 17, 2017. Data Availability Statement: All relevant data are within the paper and its Supporting Information files. Funding: Funded by Howard Hughes Medical Institute (http://www.hhmi.org): JC AR EKO. Samsung Scholarship (http://www.ssscholarship.com): JC. Mary Fieser fellowship (http://chemistry.harvard.edu): AR. DOE Grant DE-SC0012461 (https://science.energy.gov/) : YX JDR. We thank Martin Jonikas and Jonathan Weissman for sharing with us the yeast library and Matlab analysis codes; Joseph Markson for insightful discussions; members of the O'Shea laboratory for discussions; and Joseph Markson, Dennis Wykoff and Harold Kim for critical reading the manuscript. Author Contributions: Conceptualization: JC AR EKO. Data curation: JC AR YX. Formal analysis: JC. Funding acquisition: JDR EKO. Investigation: JC AR YX. Project administration: JDR EKO. Resources: JC AR YX. Software: JC. Supervision: JDR EKO. Visualization: JC. Writing – original draft: JC EKO. Writing – review & editing: JC AR YX JDR EKO. The authors have declared that no competing interests exist.Attached Files
Published - journal.pone.0176085.pdf
Supplemental Material - journal.pone.0176085.s001.tif
Supplemental Material - journal.pone.0176085.s002.xlsx
Supplemental Material - journal.pone.0176085.s003.xlsx
Supplemental Material - journal.pone.0176085.s004.xlsx
Supplemental Material - journal.pone.0176085.s005.xlsx
Supplemental Material - journal.pone.0176085.s006.xlsx
Supplemental Material - journal.pone.0176085.s007.xlsx
Files
Name | Size | Download all |
---|---|---|
md5:e6810836dda1fce5c803f3899d9795b1
|
5.2 MB | Download |
md5:8174d5030a93c9a30fcbbcfed4bce587
|
15.1 kB | Download |
md5:8295c4373d0c199d4b50ab19b145b82f
|
120.3 kB | Download |
md5:3f00acbb9033faf52401dd0c7abb2ef5
|
95.4 kB | Download |
md5:f50d00f1aae3dca246c09168ce502c19
|
51.1 kB | Download |
md5:742b13a6a2aca60d3837a87e0be4e517
|
1.6 MB | Preview Download |
md5:c75ea5b99d15163db6b35ca09e440791
|
128.9 kB | Preview Download |
md5:d66987ccb8b851633932476387a36ff5
|
760.0 kB | Download |
Additional details
- PMCID
- PMC5435139
- Eprint ID
- 78054
- Resolver ID
- CaltechAUTHORS:20170609-103921506
- Howard Hughes Medical Institute (HHMI)
- Samsung Scholarship
- Mary Fieser Fellowship
- DE-SC0012461
- Department of Energy (DOE)
- Created
-
2017-06-09Created from EPrint's datestamp field
- Updated
-
2023-06-01Created from EPrint's last_modified field