Exposure to toxic concentrations of aluminum activates a MAPK-like protein in cell suspension cultures of Coffea arabica.
Addition of a toxic concentration of aluminum (Al) to cell suspension cultures of Coffea arabica L. induced the rapid and transient activation of a protein kinase that phosphorylates myelin basic protein (MBP), as revealed by in-gel kinase assays. This enzyme with an apparent molecular mass of 58 kDa was activated shortly after cells were exposed to 50 microM AlCl(3), a concentration previously shown to produce toxicity in plant cells in vitro. The activity of this kinase dropped to basal levels after 20 min of Al addition; this activity is specific for MBP as it could not be detected when casein or histone H1 were used as substrates. Analysis of the same cell extracts with antibodies that specifically recognize bis-phosphorylated (active) mitogen-activated protein kinases (MAP kinases), revealed the presence of a phosphoprotein with an apparent molecular mass of 58 kDa, which showed the same response to Al as the protein kinase revealed by the in-gel kinase assays. Furthermore, immunoprecipitation with an antibody directed against mammalian MAP kinases depleted both the enzymatic activity and the phosphoprotein from the cell extracts, suggesting that the 58 kDa kinase and the 58 kDa phosphoprotein from C. arabica cells are the same protein, and that it can be actually a member of the MAP kinase family of protein kinases. Since its activity is enhanced dramatically after addition of AlCl(3) to the medium, we can speculate that Al toxicity in plants could be perceived through the MAP kinase signal transduction pathway.
Arroyo-Serralta
Gabriela A
GA
Unidad de Bioquímica y Biología Molecular de Plantas, Centro de Investigación Científica de Yucatán, Mérida 97200, Yucatán, México. zuniga@cicy.mx
Kú-González
Angela
A
Hernández-Sotomayor
S M Teresa
SM
Zúñiga Aguilar
José J
JJ
D000535:Aluminum / Q000633:toxicity*; D000595:Amino Acid Sequence; D002364:Caseins / Q000378:metabolism; D002478:Cells, Cultured; D040503:Coffea / Q000187:drug effects / Q000378:metabolism*; D004789:Enzyme Activation / Q000187:drug effects / Q000502:physiology; D006657:Histones / Q000378:metabolism; D020935:MAP Kinase Signaling System / Q000187:drug effects / Q000502:physiology*; D008969:Molecular Sequence Data; D004676:Myelin Basic Protein / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D010940:Plant Proteins / Q000378:metabolism*
D002364:Caseins; D006657:Histones; D004676:Myelin Basic Protein; D010940:Plant Proteins; D000535:Aluminum