Chemogenetic profiling reveals PP2A-independent cytotoxicity of proposed PP2A activators iHAP1 and DT-061
Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
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Chemogenetic profiling reveals PP2A-independent cytotoxicity of proposed PP2A activators iHAP1 and DT-061. / Vit, Gianmatteo; Duro, Joana; Rajendraprasad, Girish; Hertz, Emil P.T.; Holland, Lya Katrine Kauffeldt; Weisser, Melanie Bianca; McEwan, Brennan C.; Lopez-Mendez, Blanca; Sotelo-Parrilla, Paula; Jeyaprakash, A. Arockia; Montoya, Guillermo; Mailand, Niels; Maeda, Kenji; Kettenbach, Arminja; Barisic, Marin; Nilsson, Jakob.
I: EMBO Journal, Bind 41, Nr. 14, e110611, 2022.Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
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TY - JOUR
T1 - Chemogenetic profiling reveals PP2A-independent cytotoxicity of proposed PP2A activators iHAP1 and DT-061
AU - Vit, Gianmatteo
AU - Duro, Joana
AU - Rajendraprasad, Girish
AU - Hertz, Emil P.T.
AU - Holland, Lya Katrine Kauffeldt
AU - Weisser, Melanie Bianca
AU - McEwan, Brennan C.
AU - Lopez-Mendez, Blanca
AU - Sotelo-Parrilla, Paula
AU - Jeyaprakash, A. Arockia
AU - Montoya, Guillermo
AU - Mailand, Niels
AU - Maeda, Kenji
AU - Kettenbach, Arminja
AU - Barisic, Marin
AU - Nilsson, Jakob
N1 - Publisher Copyright: © 2022 The Authors. Published under the terms of the CC BY 4.0 license.
PY - 2022
Y1 - 2022
N2 - Protein phosphatase 2A (PP2A) is an abundant phosphoprotein phosphatase that acts as a tumor suppressor. For this reason, compounds able to activate PP2A are attractive anticancer agents. The compounds iHAP1 and DT-061 have recently been reported to selectively stabilize specific PP2A-B56 complexes to mediate cell killing. We were unable to detect direct effects of iHAP1 and DT-061 on PP2A-B56 activity in biochemical assays and composition of holoenzymes. Therefore, we undertook genome-wide CRISPR-Cas9 synthetic lethality screens to uncover biological pathways affected by these compounds. We found that knockout of mitotic regulators is synthetic lethal with iHAP1 while knockout of endoplasmic reticulum (ER) and Golgi components is synthetic lethal with DT-061. Indeed we showed that iHAP1 directly blocks microtubule assembly both in vitro and in vivo and thus acts as a microtubule poison. In contrast, DT-061 disrupts both the Golgi apparatus and the ER and lipid synthesis associated with these structures. Our work provides insight into the biological pathways perturbed by iHAP1 and DT-061 causing cellular toxicity and argues that these compounds cannot be used for dissecting PP2A-B56 biology.
AB - Protein phosphatase 2A (PP2A) is an abundant phosphoprotein phosphatase that acts as a tumor suppressor. For this reason, compounds able to activate PP2A are attractive anticancer agents. The compounds iHAP1 and DT-061 have recently been reported to selectively stabilize specific PP2A-B56 complexes to mediate cell killing. We were unable to detect direct effects of iHAP1 and DT-061 on PP2A-B56 activity in biochemical assays and composition of holoenzymes. Therefore, we undertook genome-wide CRISPR-Cas9 synthetic lethality screens to uncover biological pathways affected by these compounds. We found that knockout of mitotic regulators is synthetic lethal with iHAP1 while knockout of endoplasmic reticulum (ER) and Golgi components is synthetic lethal with DT-061. Indeed we showed that iHAP1 directly blocks microtubule assembly both in vitro and in vivo and thus acts as a microtubule poison. In contrast, DT-061 disrupts both the Golgi apparatus and the ER and lipid synthesis associated with these structures. Our work provides insight into the biological pathways perturbed by iHAP1 and DT-061 causing cellular toxicity and argues that these compounds cannot be used for dissecting PP2A-B56 biology.
KW - chemogenic profiling
KW - DT-061
KW - iHAP1
KW - phosphatase
KW - PP2A
U2 - 10.15252/embj.2022110611
DO - 10.15252/embj.2022110611
M3 - Journal article
C2 - 35695070
AN - SCOPUS:85131746365
VL - 41
JO - E M B O Journal
JF - E M B O Journal
SN - 0261-4189
IS - 14
M1 - e110611
ER -
ID: 310966538