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Protein Phosphorylation during Coconut Zygotic Embryo Development1

Ignacio Islas-Flores, Carlos Oropeza, and S.M. Teresa Hernández-Sotomayor*

Unidad de Biología Experimental, Centro de Investigación Científica de Yucatán, Apdo. Postal 87, Cordemex, Yucatán 97310, Mexico

Evidence was obtained on the occurrence of protein threonine, serine, and tyrosine (Tyr) kinases in developing coconut (Cocos nucifera L.) zygotic embryos, based on in vitro phosphorylation of proteins in the presence of [gamma -32P]ATP, alkaline treatment, and thin-layer chromatography analysis, which showed the presence of [32P]phosphoserine, [32P]phosphothreonine, and [32P]phosphotyrosine in [32P]-labeled protein hydrolyzates. Tyr kinase activity was further confirmed in extracts of embryos at different stages of development using antiphosphotyrosine monoclonal antibodies and the synthetic peptide derived from the amino acid sequence surrounding the phosphorylation site in pp60src (RR-SRC), which is specific for Tyr kinases. Anti-phosphotyrosine western blotting revealed a changing profile of Tyr-phosphorylated proteins during embryo development. Tyr kinase activity, as assayed using RR-SRC, also changed during embryo development, showing two peaks of activity, one during early and another during late embryo development. In addition, the use of genistein, a Tyr kinase inhibitor, diminished the ability of extracts to phosphorylate RR-SRC. Results presented here show the occurrence of threonine, serine, and Tyr kinases in developing coconut zygotic embryos, and suggest that protein phosphorylation, and the possible inference of Tyr phosphorylation in particular, may play a role in the coordination of the development of embryos in this species.

1   This work was supported by Fogarty International Research Collaboration Award (grant no. RO3TW00263), the Consejo Nacional de Ciencia y Tecnología (CONACYT) (grant no. 0014P-N9506), the Commission of the European Communities EC-STD (grant no. ERBTS3*CT940298), and a CONACYT Fellowship to I.I.-F. (no. 89535).
*   Corresponding author; e-mail ths{at}cicy.cicy.mx; fax 1-91-81-3900.

Plant Physiol. (1998) 118: 257-263
Copyright Clearance Center:   0032-0889/98/118//07
© 1998 American Society of Plant Physiologists




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