Molecular characterization of a cytokinin-inducible periwinkle protein showing sequence homology with pathogenesis-related proteins and the Bet v 1 allergen family.
Cytokinin treatment of periwinkle callus cultures increased the accumulation of a protein, designated T1, in two-dimensional separated protein extracts. The first 30 NH2-terminal amino acids were determined by Edman degradation and showed significant sequence homology with intracellular pathogenesis-related (IPR) plant proteins and the Bet v 1 allergen family. The deduced amino acid sequence of cDNAs coding for T1, isolated by RT-PCR and 5' RACE-PCR, exhibited an average sequence identity of 40% with both IPR and Bet v 1-related allergens. T1 and all related proteins contained a p-loop motif typically found in nucleotide-binding proteins as the most conserved sequence feature. Northern blot analysis showed that cytokinin treatment of periwinkle callus induced T1 transcripts, whereas addition of 2,4-dichlorophenoxyacetic acid inhibited this accumulation. Hybridization of genomic periwinkle DNA with the T1 cDNA suggested that the protein is encoded by a single-copy gene. Immunoblot studies with a panel of Bet v 1-specific antibodies and sera from Bet v 1 allergic individuals identified T1 as a protein that is immunologically distinct from the Bet v 1 allergen family and has no allergenic properties.
Carpin
S
S
Laboratoire de Biologie Moléculaire et Biochimie Végétale, Faculté de Pharmacie, Tours, France.
Laffer
S
S
Schoentgen
F
F
Valenta
R
R
Chénieux
J C
JC
Rideau
M
M
Hamdi
S
S
D000485:Allergens / Q000235:genetics; D000595:Amino Acid Sequence; D052179:Antigens, Plant; D001483:Base Sequence; D003583:Cytokinins / Q000494:pharmacology; D017931:DNA Primers / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D018744:DNA, Plant / Q000235:genetics; D018506:Gene Expression Regulation, Plant / Q000187:drug effects; D017343:Genes, Plant; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000096:biosynthesis / Q000235:genetics*; D010944:Plants / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D016133:Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid
D000485:Allergens; D052179:Antigens, Plant; D003583:Cytokinins; D017931:DNA Primers; D018076:DNA, Complementary; D018744:DNA, Plant; D010940:Plant Proteins; C053376:pathogenesis-related proteins, plant; C061312:Bet v 1 allergen, Betula