Gain-of-function genetic screens in human cells identify SLC transporters overcoming environmental nutrient restrictions.

Details

Serval ID
serval:BIB_ECC63CD3FA4B
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Gain-of-function genetic screens in human cells identify SLC transporters overcoming environmental nutrient restrictions.
Journal
Life science alliance
Author(s)
Rebsamen M., Girardi E., Sedlyarov V., Scorzoni S., Papakostas K., Vollert M., Konecka J., Guertl B., Klavins K., Wiedmer T., Superti-Furga G.
ISSN
2575-1077 (Electronic)
ISSN-L
2575-1077
Publication state
Published
Issued date
11/2022
Peer-reviewed
Oui
Volume
5
Number
11
Pages
e202201404
Language
english
Notes
Publication types: Journal Article
Publication Status: epublish
Abstract
Solute carrier (SLC) transporters control fluxes of nutrients and metabolites across membranes and thereby represent a critical interface between the microenvironment and cellular and subcellular metabolism. Because of substantial functional overlap, the interplay and relative contributions of SLCs in response to environmental stresses remain poorly elucidated. To infer functional relationships between SLCs and metabolites, we developed a strategy to identify SLCs able to sustain cell viability and proliferation under growth-limiting concentrations of essential nutrients. One-by-one depletion of 13 amino acids required for cell proliferation enabled gain-of-function genetic screens using a SLC-focused CRISPR/Cas9-based transcriptional activation approach to uncover transporters relieving cells from growth-limiting metabolic bottlenecks. Among the transporters identified, we characterized the cationic amino acid transporter SLC7A3 as a gene that, when up-regulated, overcame low availability of arginine and lysine by increasing their uptake, whereas SLC7A5 was able to sustain cellular fitness upon deprivation of several neutral amino acids. Moreover, we identified metabolic compensation mediated by the glutamate/aspartate transporters SLC1A2 and SLC1A3 under glutamine-limiting conditions. Overall, this gain-of-function approach using human cells uncovered functional transporter-nutrient relationships and revealed that transport activity up-regulation may be sufficient to overcome environmental metabolic restrictions.
Keywords
Amino Acid Transport Systems, Basic/genetics, Amino Acids/metabolism, Arginine, Aspartic Acid, Gain of Function Mutation, Glutamates, Glutamine, Humans, Large Neutral Amino Acid-Transporter 1, Lysine, Membrane Transport Proteins/metabolism, Nutrients
Pubmed
Open Access
Yes
Create date
27/09/2022 11:52
Last modification date
28/09/2022 5:39
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