Click chemistry–enabled CRISPR screening reveals GSK3 as a regulator of PLD signaling

Enzymes that produce second messengers are highly regulated. Revealing the mechanisms underlying such regulation is critical to understanding both how cells achieve specific signaling outcomes and return to homeostasis following a particular stimulus. Pooled genome-wide CRISPR screens are powerful u...

Full description

Saved in:
Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 2021-11, Vol.118 (48), p.1-7
Main Authors: Bumpus, Timothy W., Huang, Shiying, Tei, Reika, Baskin, Jeremy M.
Format: Article
Language:English
Subjects:
Biological Phenomena
Biological Sciences
Chemical synthesis
Click Chemistry - methods
Clustered Regularly Interspaced Short Palindromic Repeats
CRISPR
CRISPR-Cas Systems - genetics
Fluorescence
Genomes
Glycogen
Glycogen synthase kinase 3
Glycogen Synthase Kinase 3 - metabolism
Glycogen Synthase Kinase 3 - physiology
Glycogens
HEK293 Cells
Homeostasis
Humans
K562 Cells
Kinases
Lipids
Mammalian cells
Phenotypes
Phosphatidic acid
Phosphatidic Acids - metabolism
Phospholipase D
Phospholipase D - metabolism
Phospholipase D - physiology
Physical Sciences
Protein kinase C
Protein Kinase C-alpha - metabolism
Protein Kinase C-alpha - physiology
Screening
Second Messenger Systems
Second messengers
Signal Transduction
Signaling
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Enzymes that produce second messengers are highly regulated. Revealing the mechanisms underlying such regulation is critical to understanding both how cells achieve specific signaling outcomes and return to homeostasis following a particular stimulus. Pooled genome-wide CRISPR screens are powerful unbiased approaches to elucidate regulatory networks, their principal limitation being the choice of phenotype selection. Here, we merge advances in bioorthogonal fluorescent labeling and CRISPR screening technologies to discover regulators of phospholipase D (PLD) signaling, which generates the potent lipid second messenger phosphatidic acid. Our results reveal glycogen synthase kinase 3 as a positive regulator of protein kinase C and PLD signaling. More generally, this work demonstrates how bioorthogonal, activity-based fluorescent tagging can expand the power of CRISPR screening to uncover mechanisms regulating specific enzyme-driven signaling pathways in mammalian cells.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.2025265118