MAP-kinase activity in etiolated Cucumis sativus cotyledons: The effect of red and far-red light irradiation

• Published in: Plant physiology and biochemistry Vol. 63; pp. 1 - 7
• Main Authors: Alvarez-Flórez, Fagua, Vidal, Dolors, Simón, Esther
• Format: Journal Article
• Language: English
• Published: Paris Elsevier Masson SAS 01.02.2013; Elsevier
• Subjects: apigenin; Biological and medical sciences; Cotyledon - enzymology; Cotyledon - radiation effects; cotyledons; Cucumber (Cucumis sativus L.); Cucumis sativus; Cucumis sativus - enzymology; Cucumis sativus - radiation effects; Enzyme Activation - radiation effects; etiolation; far-red light; Fundamental and applied biological sciences. Psychology; genistein; Immunoblotting; Immunoprecipitation; irradiation; Light; MAP-kinase activity; mitogen-activated protein kinase; Mitogen-Activated Protein Kinases - metabolism; monoclonal antibodies; myelin basic protein; photoreceptors; phytochrome; Phytochromes; Plant physiology and development; Plant Proteins - metabolism; precipitin tests; Protein kinase; protein phosphorylation; Red and far-red light; signal transduction; Signal Transduction - radiation effects; tyrosine; Protein kinase; Phytochromes; Cucumber (Cucumis sativus L.); MAP-kinase activity; Red and far-red light; Cotyledon; Enzyme; Transferases; Non-specific serine/threonine protein kinase; Cucurbitaceae; Red light; Mitogen-activated protein kinase; Cucumis sativus; Etiolation; Vegetable crop; Plant physiology; Dicotyledones; Angiospermae; Phytochrome; Irradiation; Far red light; Spermatophyta
• ISSN: 0981-9428; 1873-2690
• DOI: 10.1016/j.plaphy.2012.11.008

Phytochrome (phy) signalling in plants may be transduced through protein phosphorylation. Mitogen-activated protein kinase (MAP-kinase, MAPK) activity and the effect of R (red) and FR (far-red) light irradiation on MAPK activity were studied in etiolated Cucumis sativus L. cotyledons. By in vitro protein phosphorylation and in-gel assays with myelin basic protein (MBP), a protein band (between 48 and 45 kDa) with MAPK-like activity was detected. The addition to the phosphorylation buffer of specific protein phosphatase (PTP) inhibitors (Na3VO4 and NaF) and genistein, apigenin or PD98059 as MAPK inhibitors allowed us to confirm the MAPK activity of the protein band. Irradiation of etiolated cotyledons with FR light for 5, 10 or 60 min rapidly and transiently stimulated the MAPK activity of the protein band. This suggests that there was a very low fluence response (VLFR) of phys. In addition, 15 min of R light irradiation or a sequential treatment of 15 min of R plus 5 min of FR also increased MAPK activity. The stimulatory effect of R light was also attributed to the same photoreceptor, which suggests that MAPKs are involved in phytochrome signal transduction. Protein immunoprecipitation and immunoblotting analysis with the polyclonal antibody anti-pERK1/2 (Tyr 204) and the monoclonal antibody anti-phosphotyrosine PY20 allowed us to recognize the above mentioned protein band as two proteins with molecular masses (Mr) of approximately 47 and 45 kDa, and MAPK activity. The biochemical and immunological properties showed by the proteins detected indicated that they were members of the MAPK family phosphorylated in tyrosine residues. ► Two PKs with MAPK properties were detected in etiolated cucumber cotyledons. ► FR light rapidly and transiently stimulated MAPK activity by a VLFR of phytochrome. ► The results suggest that MAPKs are involved in phytochrome signal transduction.