Depletion of WRN protein causes RACK1 to activate several protein kinase C isoforms
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Depletion of WRN protein causes RACK1 to activate several protein kinase C isoforms. / Massip, L; Garand, C; Labbé, A; Perreault, E; Turaga, R V N; Bohr, V A; Lebel, M.
I: Oncogene, Bind 29, Nr. 10, 11.03.2010, s. 1486-97.Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
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T1 - Depletion of WRN protein causes RACK1 to activate several protein kinase C isoforms
AU - Massip, L
AU - Garand, C
AU - Labbé, A
AU - Perreault, E
AU - Turaga, R V N
AU - Bohr, V A
AU - Lebel, M
PY - 2010/3/11
Y1 - 2010/3/11
N2 - Werner's syndrome (WS) is a rare autosomal disease characterized by the premature onset of several age-associated pathologies. The protein defective in patients with WS (WRN) is a helicase/exonuclease involved in DNA repair, replication, transcription and telomere maintenance. In this study, we show that a knock down of the WRN protein in normal human fibroblasts induces phosphorylation and activation of several protein kinase C (PKC) enzymes. Using a tandem affinity purification strategy, we found that WRN physically and functionally interacts with receptor for activated C-kinase 1 (RACK1), a highly conserved anchoring protein involved in various biological processes, such as cell growth and proliferation. RACK1 binds strongly to the RQC domain of WRN and weakly to its acidic repeat region. Purified RACK1 has no impact on the helicase activity of WRN, but selectively inhibits WRN exonuclease activity in vitro. Interestingly, knocking down RACK1 increased the cellular frequency of DNA breaks. Depletion of the WRN protein in return caused a fraction of nuclear RACK1 to translocate out of the nucleus to bind and activate PKCdelta and PKCbetaII in the membrane fraction of cells. In contrast, different DNA-damaging treatments known to activate PKCs did not induce RACK1/PKCs association in cells. Overall, our results indicate that a depletion of the WRN protein in normal fibroblasts causes the activation of several PKCs through translocation and association of RACK1 with such kinases.
AB - Werner's syndrome (WS) is a rare autosomal disease characterized by the premature onset of several age-associated pathologies. The protein defective in patients with WS (WRN) is a helicase/exonuclease involved in DNA repair, replication, transcription and telomere maintenance. In this study, we show that a knock down of the WRN protein in normal human fibroblasts induces phosphorylation and activation of several protein kinase C (PKC) enzymes. Using a tandem affinity purification strategy, we found that WRN physically and functionally interacts with receptor for activated C-kinase 1 (RACK1), a highly conserved anchoring protein involved in various biological processes, such as cell growth and proliferation. RACK1 binds strongly to the RQC domain of WRN and weakly to its acidic repeat region. Purified RACK1 has no impact on the helicase activity of WRN, but selectively inhibits WRN exonuclease activity in vitro. Interestingly, knocking down RACK1 increased the cellular frequency of DNA breaks. Depletion of the WRN protein in return caused a fraction of nuclear RACK1 to translocate out of the nucleus to bind and activate PKCdelta and PKCbetaII in the membrane fraction of cells. In contrast, different DNA-damaging treatments known to activate PKCs did not induce RACK1/PKCs association in cells. Overall, our results indicate that a depletion of the WRN protein in normal fibroblasts causes the activation of several PKCs through translocation and association of RACK1 with such kinases.
KW - Blotting, Western
KW - Cell Line, Tumor
KW - Cells, Cultured
KW - DNA Damage
KW - Enzyme Activation
KW - Exodeoxyribonucleases
KW - Fibroblasts
KW - GTP-Binding Proteins
KW - Humans
KW - Neoplasm Proteins
KW - Phosphoproteins
KW - Phosphorylation
KW - Protein Binding
KW - Protein Kinase C
KW - Protein Kinase C-delta
KW - RNA Interference
KW - RecQ Helicases
KW - Receptors, Cell Surface
KW - p38 Mitogen-Activated Protein Kinases
U2 - 10.1038/onc.2009.443
DO - 10.1038/onc.2009.443
M3 - Journal article
C2 - 19966859
VL - 29
SP - 1486
EP - 1497
JO - Oncogene
JF - Oncogene
SN - 0950-9232
IS - 10
ER -
ID: 33491474