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StCDPK2 expression and activity reveal a highly responsive potato calcium-dependent protein kinase involved in light signalling

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Abstract

Calcium-dependent protein kinases (CDPKs) are essential calcium sensors. In this work, we have studied StCDPK2 isoform from potato both at gene and protein level. StCdpk2 genomic sequence contains eight exons and seven introns, as was observed for StCdpk1. There is one copy of the gene per genome located in chromosome 7. StCDPK2 encodes an active CDPK of 515 aminoacids, with an apparent MW of 57 kDa, which presents myristoylation and palmitoylation consensus in its N-terminus. StCDPK2 is highly expressed in leaves and green sprouts; enhanced expression was detected under light treatment, which corresponds well with light responsive cis-acting elements found in its promoter sequence. Antibodies against the recombinant StCDPK2::6xHis protein detected this isoform in soluble and particulate fractions from leaves. StCDPK2 autophosphorylation and kinase activity are both calcium dependent reaching half maximal activation at 0.6 μM calcium. The active kinase is autophosphorylated on serine and tyrosine residues and its activity is negatively modulated by phosphatidic acid (PA). Our results reveal StCDPK2 as a signalling element involved in plant growth and development and show that its activity is tightly regulated.

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Abbreviations

ABA:

Absicic acid

CDPK:

Calcium-dependent protein kinase

DGPP:

Diacylglycerol pyrophosphate

JA:

Jasmonic acid

PA:

Phosphatidic acid

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Acknowledgments

We thank technical assistance of Karen McClean. We thank Dr. Sara Maldonado for helpful discussion and the technical assistance of Verónica Lainez. We thank Dr Salomé Prat Monguío (CSIC, Madrid, Spain) for the cDNA library from tuberizing stolons and Dr. Ana L. Villasuso and Dr. Estela E. Machado for providing the phospholipids. This work was supported by grants from CONICET, UBA, FONCYT-ANPCYT and the Royal Society of Edinburgh. RMU and PRG are members of CONICET. VG, CG and SB are fellows from CONICET.

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Author notes

  1. Pablo Rubén Gargantini

    Present address: Facultad de Medicina, Universidad Católica de Córdoba (UCC), Jacinto Ríos 571, 5000, Córdoba, Argentina

Authors and Affiliations

  1. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular (INGEBI), CONICET and Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Vuelta de Obligado 2490 2do piso, 1428, Buenos Aires, Argentina

    Verónica Giammaria, Carolina Grandellis, Sandra Bachmann, Pablo Rubén Gargantini & Rita María Ulloa

  2. Laboratorio de Agrobiotecnología, EEA Balcarce-INTA, C.C. 276 (7620), Balcarce, Argentina

    Sergio Enrique Feingold

  3. Genetics Programme, Scottish Crop Research Institute (SCRI), Dundee, DD2 5DA, UK

    Glenn Bryan

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  1. Verónica Giammaria
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  2. Carolina Grandellis
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  7. Rita María Ulloa
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Correspondence to Rita María Ulloa.

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Giammaria, V., Grandellis, C., Bachmann, S. et al. StCDPK2 expression and activity reveal a highly responsive potato calcium-dependent protein kinase involved in light signalling. Planta 233, 593–609 (2011). https://doi.org/10.1007/s00425-010-1319-2

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