Two expressed soybean genes with high sequence identity to tomato Pti1 kinase lack autophosphorylation activity.
An important signaling pathway for disease resistance in tomato involves the R gene product Pto which phosphorylates Ptil, a downstream member of this signaling cascade. Both Pto and Pti1 are Ser/Thr protein kinases capable of autophosphorylation in vitro. Two soybean (Glycine max L. Merr. var. Hobbit) cDNAs (sPti1a and sPti1b) were cloned and sequenced and found to each have 78% amino acid sequence identity with tomato Pti1. Glutathione S-transferase fusions of sPti1a and b expressed in Escherichia coli did not autophosphorylate in vitro, but were efficiently phosphorylated by tomato Pto. Replacement of Tyr197 with an Asp that is invariant at this position in other protein kinases did not restore autophosphorylation activity to sPti1a or b. Tyr197 was also present in the Pti1 homologues of three distant relatives of G. max. Together these results suggest that soybean Pti1 might function in a Pto-like signaling pathway that does not require Pti1 kinase activity.
Staswick
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Department of Agronomy, University of Nebraska-Lincoln, 68583-0915, USA. pstaswick1@unl.edu
D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000378:metabolism; D004926:Escherichia coli / Q000378:metabolism; D017343:Genes, Plant; D005982:Glutathione Transferase / Q000378:metabolism; D007113:Immunity, Innate; D018551:Solanum lycopersicum / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics; D008969:Molecular Sequence Data; D005810:Multigene Family; D010766:Phosphorylation; D010940:Plant Proteins / Q000737:chemistry* / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid; D015398:Signal Transduction; D013025:Glycine max / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics
D018076:DNA, Complementary; D010940:Plant Proteins; D005982:Glutathione Transferase; D017346:Protein Serine-Threonine Kinases