Identification and characterization of novel components of a Ca 2+/calmodulin-dependent protein kinase cascade in HeLa cells

Yumi Ishikawa; Hiroshi Tokumitsu; Hiroyuki Inuzuka; Maki Murata-Hori; Hiroshi Hosoya; Ryoji Kobayashi

doi:10.1016/S0014-5793(03)00817-2

Identification and characterization of novel components of a Ca ²⁺/calmodulin-dependent protein kinase cascade in HeLa cells

Yumi Ishikawa, Hiroshi Tokumitsu, Hiroyuki Inuzuka, Maki Murata-Hori, Hiroshi Hosoya, Ryoji Kobayashi

Research output: Contribution to journal › Article › peer-review

30 Citations (Scopus)

Abstract

In this report, we cloned a novel calmodulin-kinase (CaM-KIδ) from HeLa cells and characterized its activation mechanism. CaM-KIδ exhibits Ca²⁺/CaM-dependent activity that is enhanced (∼30-fold) in vitro by phosphorylation of its Thr180 by CaM-K kinase (CaM-KK)α, consistent with detection of CaM-KIδ-activating activity in HeLa cells. We also identified a novel CaM-KKβ isoform (CaM-KKβ-3) in HeLa cells whose activity was highly Ca²⁺/CaM-independent. Transiently expressed CaM-KIδ exhibited enhanced protein kinase activity in HeLa cells without ionomycin stimulation. This sustained activation of CaM-KIδ was completely abolished by Thr180Ala mutation and inhibited by CaM-KK inhibitor, STO-609, indicating a functional CaM-KK/CaM-KIδ cascade in HeLa cells.

Original language	English
Pages (from-to)	57-63
Number of pages	7
Journal	FEBS Letters
Volume	550
Issue number	1-3
DOIs	https://doi.org/10.1016/S0014-5793(03)00817-2
Publication status	Published - Aug 28 2003
Externally published	Yes

Keywords

Ca
CaM-K kinase
CaM-kinase cascade
CaM-kinaseδ

ASJC Scopus subject areas

Biophysics
Structural Biology
Biochemistry
Molecular Biology
Genetics
Cell Biology

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10.1016/S0014-5793(03)00817-2

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title = "Identification and characterization of novel components of a Ca 2+/calmodulin-dependent protein kinase cascade in HeLa cells",

abstract = "In this report, we cloned a novel calmodulin-kinase (CaM-KIδ) from HeLa cells and characterized its activation mechanism. CaM-KIδ exhibits Ca2+/CaM-dependent activity that is enhanced (∼30-fold) in vitro by phosphorylation of its Thr180 by CaM-K kinase (CaM-KK)α, consistent with detection of CaM-KIδ-activating activity in HeLa cells. We also identified a novel CaM-KKβ isoform (CaM-KKβ-3) in HeLa cells whose activity was highly Ca2+/CaM-independent. Transiently expressed CaM-KIδ exhibited enhanced protein kinase activity in HeLa cells without ionomycin stimulation. This sustained activation of CaM-KIδ was completely abolished by Thr180Ala mutation and inhibited by CaM-KK inhibitor, STO-609, indicating a functional CaM-KK/CaM-KIδ cascade in HeLa cells.",

keywords = "Ca, CaM-K kinase, CaM-kinase cascade, CaM-kinaseδ",

author = "Yumi Ishikawa and Hiroshi Tokumitsu and Hiroyuki Inuzuka and Maki Murata-Hori and Hiroshi Hosoya and Ryoji Kobayashi",

year = "2003",

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T1 - Identification and characterization of novel components of a Ca 2+/calmodulin-dependent protein kinase cascade in HeLa cells

AU - Ishikawa, Yumi

AU - Tokumitsu, Hiroshi

AU - Inuzuka, Hiroyuki

AU - Murata-Hori, Maki

AU - Hosoya, Hiroshi

AU - Kobayashi, Ryoji

PY - 2003/8/28

Y1 - 2003/8/28

N2 - In this report, we cloned a novel calmodulin-kinase (CaM-KIδ) from HeLa cells and characterized its activation mechanism. CaM-KIδ exhibits Ca2+/CaM-dependent activity that is enhanced (∼30-fold) in vitro by phosphorylation of its Thr180 by CaM-K kinase (CaM-KK)α, consistent with detection of CaM-KIδ-activating activity in HeLa cells. We also identified a novel CaM-KKβ isoform (CaM-KKβ-3) in HeLa cells whose activity was highly Ca2+/CaM-independent. Transiently expressed CaM-KIδ exhibited enhanced protein kinase activity in HeLa cells without ionomycin stimulation. This sustained activation of CaM-KIδ was completely abolished by Thr180Ala mutation and inhibited by CaM-KK inhibitor, STO-609, indicating a functional CaM-KK/CaM-KIδ cascade in HeLa cells.

AB - In this report, we cloned a novel calmodulin-kinase (CaM-KIδ) from HeLa cells and characterized its activation mechanism. CaM-KIδ exhibits Ca2+/CaM-dependent activity that is enhanced (∼30-fold) in vitro by phosphorylation of its Thr180 by CaM-K kinase (CaM-KK)α, consistent with detection of CaM-KIδ-activating activity in HeLa cells. We also identified a novel CaM-KKβ isoform (CaM-KKβ-3) in HeLa cells whose activity was highly Ca2+/CaM-independent. Transiently expressed CaM-KIδ exhibited enhanced protein kinase activity in HeLa cells without ionomycin stimulation. This sustained activation of CaM-KIδ was completely abolished by Thr180Ala mutation and inhibited by CaM-KK inhibitor, STO-609, indicating a functional CaM-KK/CaM-KIδ cascade in HeLa cells.

KW - Ca

KW - CaM-K kinase

KW - CaM-kinase cascade

KW - CaM-kinaseδ

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