A genome-wide siRNA screen in mammalian cells for regulators of S6 phosphorylation

PloS One
Angela PapageorgiouJoseph Avruch

Abstract

mTOR complex1, the major regulator of mRNA translation in all eukaryotic cells, is strongly activated in most cancers. We performed a genome-wide RNAi screen in a human cancer cell line, seeking genes that regulate S6 phosphorylation, readout of mTORC1 activity. Applying a stringent selection, we retrieved nearly 600 genes wherein at least two RNAis gave significant reduction in S6-P. This cohort contains known regulators of mTOR complex 1 and is significantly enriched in genes whose depletion affects the proliferation/viability of the large set of cancer cell lines in the Achilles database in a manner paralleling that caused by mTOR depletion. We next examined the effect of RNAi pools directed at 534 of these gene products on S6-P in TSC1 null mouse embryo fibroblasts. 76 RNAis reduced S6 phosphorylation significantly in 2 or 3 replicates. Surprisingly, among this cohort of genes the only elements previously associated with the maintenance of mTORC1 activity are two subunits of the vacuolar ATPase and the CUL4 subunit DDB1. RNAi against a second set of 84 targets reduced S6-P in only one of three replicates. However, an indication that this group also bears attention is the presence of rpS6KB1 itself, Rac1 and MAP4K3, a protei...Continue Reading

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Citations

Mar 13, 2016·Autoimmunity Reviews·Andreas V Goules, Athanasios G Tzioufas
Oct 7, 2015·Frontiers in Genetics·Alexander Agrotis, Robin Ketteler
May 28, 2017·Applied and Environmental Microbiology·Pak Yang ChumChristopher P Landowski
Jan 30, 2018·Biotechnology Journal·Ricardo Valdés-Bango Curell, Niall Barron
Aug 10, 2017·Applied Microbiology and Biotechnology·Chit TamTzi Bun Ng
Apr 5, 2020·Nature Communications·Nikolaus A WatsonJonathan M G Higgins

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Methods Mentioned

BETA
GTPase
transfection
ubiquitination
transfectability
GTPases
FCS

Software Mentioned

Coasy
panther
MetaXpress
TIBCO
Genego
Spotfire DecisionSite
spotfire

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