The impact of calmodulin on the cell cycle analyzed in a novel human cellular genetic system
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The impact of calmodulin on the cell cycle analyzed in a novel human cellular genetic system. / Munk, Mads; Alcalde, Juan; Lorentzen, Lasse; Villalobo, Antonio; Berchtold, Martin W.; Panina, Svetlana.
I: Cell Calcium, Bind 88, 102207, 2020.Publikation: Bidrag til tidsskrift › Tidsskriftartikel › fagfællebedømt
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T1 - The impact of calmodulin on the cell cycle analyzed in a novel human cellular genetic system
AU - Munk, Mads
AU - Alcalde, Juan
AU - Lorentzen, Lasse
AU - Villalobo, Antonio
AU - Berchtold, Martin W.
AU - Panina, Svetlana
PY - 2020
Y1 - 2020
N2 - Calmodulin (CaM) is the principle mediator of the Ca2+ signal in all eukaryotic cells. A huge variety of basic cellular processes including cell cycle control, proliferation, secretion and motility, among many others are governed by CaM, which regulates activities of myriads of target proteins. Mammalian CaM is encoded by three genes localized on different chromosomes all producing an identical protein. In this study, we have generated HeLa human cancer cells conditionally expressing CaM in a genetic background with all three genes inactivated by CRISPR/Cas9. We demonstrate that downregulation of ectopically expressed CaM is achieved after 120 h, when cells are arrested in the M phase of the cell cycle. We show for the first time that CaM downregulation in human cancer cells is followed by a multinucleated senescent state as indicated by expression of β-galactosidase as well as cell morphology typical for senescent cells. Our newly generated genetic system may be useful for the analysis of other CaM regulated processes in eukaryotic cells in the absence of endogenous CaM genes.
AB - Calmodulin (CaM) is the principle mediator of the Ca2+ signal in all eukaryotic cells. A huge variety of basic cellular processes including cell cycle control, proliferation, secretion and motility, among many others are governed by CaM, which regulates activities of myriads of target proteins. Mammalian CaM is encoded by three genes localized on different chromosomes all producing an identical protein. In this study, we have generated HeLa human cancer cells conditionally expressing CaM in a genetic background with all three genes inactivated by CRISPR/Cas9. We demonstrate that downregulation of ectopically expressed CaM is achieved after 120 h, when cells are arrested in the M phase of the cell cycle. We show for the first time that CaM downregulation in human cancer cells is followed by a multinucleated senescent state as indicated by expression of β-galactosidase as well as cell morphology typical for senescent cells. Our newly generated genetic system may be useful for the analysis of other CaM regulated processes in eukaryotic cells in the absence of endogenous CaM genes.
KW - Calmodulin
KW - Cell cycle
KW - CRISPR/Cas9
KW - Mitotic arrest
KW - Senescence
KW - Tet-Off system
U2 - 10.1016/j.ceca.2020.102207
DO - 10.1016/j.ceca.2020.102207
M3 - Journal article
C2 - 32408024
AN - SCOPUS:85084378839
VL - 88
JO - Cell Calcium
JF - Cell Calcium
SN - 0143-4160
M1 - 102207
ER -
ID: 244573348