Arabidopsis homologs of the shaggy and GSK-3 protein kinases: molecular cloning and functional expression in Escherichia coli.
The conservation in evolution of fundamental signal transduction modules offers a means of isolating genes likely to be involved in plant development. We have amplified by PCR Arabidopsis cDNA and genomic sequences related to the product of the shaggy/zeste-white 3 (sgg) segment polarity gene of Drosophila. This regulatory protein is functionally homologous to glycogen synthase kinase-3 in mammals (GSK-3), which regulates, among others, the DNA-binding activity of the c-jun/AP1 transcription factor. Analysis of PCR products led to the identification of five genes; for two of which, corresponding full-length cDNAs, ASK-alpha and gamma (for Arabidopsis shaggy-related protein kinase), were characterized. The encoded proteins were 70% identical to GSK-3 and sgg over the protein kinase catalytic domain and, after production in Escherichia coli, autophosphorylated mainly on threonine and serine residues, but phosphotyrosine was also detected. ASK-alpha and ASK-gamma also phosphorylated phosphatase inhibitor-2 and myelin basic protein, on threonine and serine, respectively. The high conservation of the protein kinases of GSK-3 family, and their action at the transcriptional level, suggest that the ASK proteins have important functions in higher plants.
Bianchi
M W
MW
Centre de Recherches sur les Plantes URA 1128, Université de Paris-Sud, Orsay, France.
Guivarc'h
D
D
Thomas
M
M
Woodgett
J R
JR
Kreis
M
M
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000378:metabolism; D000818:Animals; D017360:Arabidopsis / Q000201:enzymology / Q000235:genetics*; D029681:Arabidopsis Proteins; D001483:Base Sequence; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000235:genetics*; D003001:Cloning, Molecular; D018076:DNA, Complementary; D004330:Drosophila / Q000235:genetics; D029721:Drosophila Proteins; D004926:Escherichia coli; D017343:Genes, Plant; D038362:Glycogen Synthase Kinase 3; D038341:Glycogen Synthase Kinases; D008869:Microtubule-Associated Proteins / Q000235:genetics*; D008969:Molecular Sequence Data; D004676:Myelin Basic Protein / Q000378:metabolism; D010766:Phosphorylation; D010940:Plant Proteins / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011494:Protein Kinases / Q000235:genetics*; D011505:Protein-Tyrosine Kinases / Q000235:genetics; D011506:Proteins / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins; D017386:Sequence Homology, Amino Acid; D000097847:Protein Phosphatase Inhibitory Proteins
D000596:Amino Acids; D029681:Arabidopsis Proteins; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D018076:DNA, Complementary; D029721:Drosophila Proteins; D038362:Glycogen Synthase Kinase 3; D038341:Glycogen Synthase Kinases; D008869:Microtubule-Associated Proteins; D004676:Myelin Basic Protein; D010940:Plant Proteins; D011494:Protein Kinases; D011505:Protein-Tyrosine Kinases; D011506:Proteins; D029701:Saccharomyces cerevisiae Proteins; C024833:protein phosphatase inhibitor-2; D000097847:Protein Phosphatase Inhibitory Proteins; C085937:ATSK12 protein, Arabidopsis; C080335:Sgg protein, Drosophila; C085936:ATSK11 protein, Arabidopsis; C068124:MCK1 protein, S cerevisiae