tailieunhanh - Báo cáo khoa học: Cloning and characterization of CBL-CIPK signalling components from a legume (Pisum sativum)

The studies on calcium sensor calcineurin B-like protein (CBL) and CBL interacting protein kinases (CIPK) are limited to Arabidopsisand rice and their functional role is only beginning to emerge. Here, we present cloning and characterization of a protein kinase (PsCIPK) from a legume, pea, with novel properties. | ềFEBS Journal Cloning and characterization of CBL-CIPK signalling components from a legume Pisum sativum Shilpi Mahajan Sudhir K. Sopory and Narendra Tuteja Plant Molecular Biology InternationalCentre for Genetic Engineering and Biotechnology New Delhi India Keywords abscisic acid abiotic stress biotic stress calcium sensor CBL CIPK Correspondence N. Tuteja Plant Molecular Biology InternationalCentre for Genetic Engineering and Biotechnology Aruna Asaf Ali Marg New Delhi 110067 India Fax 91 11 26162316 E-mail narendra@ Note The sequences reported in this paper have been deposited in the General Bank database accession nos. AY134619 pea CBL cDNA AY883569 pea CBL genomic clone AY191840 pea CIPK cDnA . Received 7 October 2005 revised 11 December 2005 accepted 19 December 2005 doi The studies on calcium sensor calcineurin B-like protein CBL and CBL interacting protein kinases CIPK are limited to Arabidopsis and rice and their functional role is only beginning to emerge. Here we present cloning and characterization of a protein kinase PsCIPK from a legume pea with novel properties. The PsCIPK gene is intronless and encodes a protein that showed partial homology to the members of CIPK family. The recombinant PsCIPK protein was autophosphorylated at Thr residue s . Immunoprecipitation and yeast two-hybrid analysis showed direct interaction of PsCIPK with PsCBL whose cDNA and genomic DNA were also cloned in this study. PsCBL showed homology to AtCBL3 and contained calcium-binding activity. We demonstrate for the first time that PsCBL is phosphorylated at its Thr residue s by PsCIPK. Immunofluorescence confocal microscopy showed that PsCBL is exclusively localized in the cytosol whereas PsCIPK is localized in the cytosol and the outer membrane. The exposure of plants to NaCl cold and wounding co-ordinately upregulated the expression of PsCBL and PsCIPK genes. The transcript levels of both genes were also coordinately stimulated .