The myristoylated amino-terminus of an Arabidopsis calcium-dependent protein kinase mediates plasma membrane localization

The myristoylated amino-terminus of an Arabidopsis calcium-dependent protein kinase mediates... Calcium-dependent protein kinases (CDPK) are a major group of calcium-stimulated kinases found in plants and some protists. Many CDPKs are membrane-associated, presumably because of lipid modifications at their amino termini. We investigated the subcellular location and myristoylation of AtCPK5, a member of the Arabidopsis CDPK family. Most AtCPK5 was associated with the plasma membrane as demonstrated by two-phase fractionation of plant microsomes and by in vivo detection of AtCPK5-GFP fusion proteins. AtCPK5 was a substrate for plant N-myristoyltransferase and myristoylation was prevented by converting the glycine at the proposed site of myristate attachment to alanine (G2A). In transgenic plants, a G2A mutation completely abolished AtCPK5 membrane association, indicating that myristoylation was essential for membrane binding. The first sixteen amino acids of AtCPK5 were sufficient to direct plasma membrane localization. In addition, differentially phosphorylated forms of AtCPK5 were detected both in planta and after expression of AtCPK5 in a cell-free plant extract. Our results demonstrate that AtCPK5 is myristoylated at its amino terminus and that myristoylation is required for membrane binding. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Plant Molecular Biology Springer Journals

The myristoylated amino-terminus of an Arabidopsis calcium-dependent protein kinase mediates plasma membrane localization

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Publisher
Springer Journals
Copyright
Copyright © 2013 by The Author(s)
Subject
Life Sciences; Plant Sciences; Biochemistry, general; Plant Pathology
ISSN
0167-4412
eISSN
1573-5028
D.O.I.
10.1007/s11103-013-0061-0
Publisher site
See Article on Publisher Site

Abstract

Calcium-dependent protein kinases (CDPK) are a major group of calcium-stimulated kinases found in plants and some protists. Many CDPKs are membrane-associated, presumably because of lipid modifications at their amino termini. We investigated the subcellular location and myristoylation of AtCPK5, a member of the Arabidopsis CDPK family. Most AtCPK5 was associated with the plasma membrane as demonstrated by two-phase fractionation of plant microsomes and by in vivo detection of AtCPK5-GFP fusion proteins. AtCPK5 was a substrate for plant N-myristoyltransferase and myristoylation was prevented by converting the glycine at the proposed site of myristate attachment to alanine (G2A). In transgenic plants, a G2A mutation completely abolished AtCPK5 membrane association, indicating that myristoylation was essential for membrane binding. The first sixteen amino acids of AtCPK5 were sufficient to direct plasma membrane localization. In addition, differentially phosphorylated forms of AtCPK5 were detected both in planta and after expression of AtCPK5 in a cell-free plant extract. Our results demonstrate that AtCPK5 is myristoylated at its amino terminus and that myristoylation is required for membrane binding.

Journal

Plant Molecular BiologySpringer Journals

Published: Apr 23, 2013

References

  • OsCDPK13, a calcium-dependent protein kinase gene from rice, is induced by cold and gibberellin in rice leaf sheath
    Abbasi, F; Onodera, H; Toki, S; Tanaka, H; Komatsu, S
  • Rice SPK, a calmodulin-like domain protein kinase, is required for storage product accumulation during seed development: phosphorylation of sucrose synthase is a possible factor
    Asano, T; Kunieda, N; Omura, Y; Ibe, H; Kawasaki, T; Takano, M; Sato, M; Furuhashi, H; Mujin, T; Takaiwa, F; Wu Cy, CY; Tada, Y; Satozawa, T; Sakamoto, M; Shimada, H
  • Calcium signaling through protein kinases. The Arabidopsis calcium-dependent protein kinase gene family
    Cheng, S; Willmann, MR; Chen, H; Sheen, J
  • Arabidopsis calcium-dependent protein kinase AtCPK32 interacts with ABF4, a transcriptional regulator of abscisic acid-responsive gene expression, and modulates its activity
    Choi, HI; Park, HJ; Park, JH; Kim, S; Im, MY; Seo, HH; Kim, YW; Hwang, I; Kim, SY
  • Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana
    Clough, SJ; Bent, AF
  • AtCPK1 calcium-dependent protein kinase mediates pathogen resistance in Arabidopsis
    Coca, M; San Segundo, B
  • Soluble, highly fluorescent variants of green fluorescent protein (GFP) for use in higher plants
    Davis, SJ; Vierstra, RD
  • Immunocytological localization of an epitope-tagged plasma membrane proton pump (H+-ATPase) in phloem companion cells
    DeWitt, ND; Sussman, MR
  • Cloning, expression and N-terminal myristoylation of CpCPK1, a calcium-dependent protein kinase from zucchini (Cucurbita pepo L.)
    Ellard-Ivey, M; Hopkins, RB; White, T; Lomax, TL

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