A comprehensive siRNA screen for kinases that suppress macroautophagy in optimal growth conditions
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A comprehensive siRNA screen for kinases that suppress macroautophagy in optimal growth conditions. / Szyniarowski, Piotr; Corcelle-Termeau, Elisabeth; Farkas, Thomas; Høyer-Hansen, Maria; Nylandsted, Jesper; Kallunki, Tuula; Jaattela, Marja.
I: Autophagy, Bind 7, Nr. 8, 2011, s. 892-903.Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
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T1 - A comprehensive siRNA screen for kinases that suppress macroautophagy in optimal growth conditions
AU - Szyniarowski, Piotr
AU - Corcelle-Termeau, Elisabeth
AU - Farkas, Thomas
AU - Høyer-Hansen, Maria
AU - Nylandsted, Jesper
AU - Kallunki, Tuula
AU - Jaattela, Marja
PY - 2011
Y1 - 2011
N2 - Macroautophagy is a catabolic process that maintains cellular homeostasis and protects cells against various external stresses including starvation. Except for the identification of the Akt-mTORC1 pathway as a major negative regulator, little is known about signaling networks that control macroautophagy under optimal growth conditions. Therefore, we screened a human kinome siRNA library for siRNAs that increase the number of autophagosomes in normally growing MCF-7 human breast carcinoma cells, and identified 10 kinases as regulators of constitutive macroautophagy. Further analysis of these kinases with respect to the autophagic flux, kinase signaling and endolysosomal function identified WNK2 as a positive regulator of autophagosome maturation and nine others as macroautophagy inhibitors. The depletion of MK2, PACSIN1, DAPK2, CDKL3 and SCYL1 functioned upstream of Akt-mTORC1 pathway, whereas CSNK1A1, BUB1, PKLR and NEK4 suppressed autophagosome formation downstream or independent of mTORC1. Importantly, all identified kinases except for BUB1 regulated macroautophagy also in immortalized MCF-10A breast epithelial cells. The kinases identified here shed light to the complex regulation of macroautophagy and open new possibilities for its pharmacological manipulation.
AB - Macroautophagy is a catabolic process that maintains cellular homeostasis and protects cells against various external stresses including starvation. Except for the identification of the Akt-mTORC1 pathway as a major negative regulator, little is known about signaling networks that control macroautophagy under optimal growth conditions. Therefore, we screened a human kinome siRNA library for siRNAs that increase the number of autophagosomes in normally growing MCF-7 human breast carcinoma cells, and identified 10 kinases as regulators of constitutive macroautophagy. Further analysis of these kinases with respect to the autophagic flux, kinase signaling and endolysosomal function identified WNK2 as a positive regulator of autophagosome maturation and nine others as macroautophagy inhibitors. The depletion of MK2, PACSIN1, DAPK2, CDKL3 and SCYL1 functioned upstream of Akt-mTORC1 pathway, whereas CSNK1A1, BUB1, PKLR and NEK4 suppressed autophagosome formation downstream or independent of mTORC1. Importantly, all identified kinases except for BUB1 regulated macroautophagy also in immortalized MCF-10A breast epithelial cells. The kinases identified here shed light to the complex regulation of macroautophagy and open new possibilities for its pharmacological manipulation.
KW - Autophagy
KW - Breast
KW - Cell Line, Tumor
KW - Cell Proliferation
KW - Endocytosis
KW - Epithelial Cells
KW - Female
KW - Genetic Testing
KW - Humans
KW - Lysosomes
KW - Microtubule-Associated Proteins
KW - Phosphatidylinositol 3-Kinases
KW - Phosphotransferases
KW - Protein Transport
KW - Proteins
KW - Proto-Oncogene Proteins c-akt
KW - RNA, Small Interfering
KW - Signal Transduction
M3 - Journal article
C2 - 21508686
VL - 7
SP - 892
EP - 903
JO - Autophagy
JF - Autophagy
SN - 1554-8627
IS - 8
ER -
ID: 38488675