The defense-related rice gene Pir7b encodes an alpha/beta hydrolase fold protein exhibiting esterase activity towards naphthol AS-esters.
Acquired resistance of rice to Pyricularia oryzae, the causing agent of rice blast, can be induced by inoculation with the non-host pathogen Pseudomonas syringae pv. syringae. We have previously cloned a cDNA and a corresponding gene (Pir7b) whose transcripts accumulate upon infiltration with the resistance-inducing bacteria. The putative encoded product Pir7b exhibits significant sequence similarity to two recently cloned hydroxynitrile lyases from Manihot esculenta (cassava) and Hevea brasisliensis, enzymes involved in the release of hydrogen cyanide from cyanogenic glycosides. As rice does not contain cyanogenic glycosides, a similar function of Pir7b appears unplausible. In order to functionally characterize the protein, recombinant Pir7b was produced in Escherichia coli and Saccharomyces cerevisiae. We show that recombinant Pir7b does not have hydroxynitrile lyase activity, but exhibits esterase activity towards naphthol AS-acetate. Using Pir7b-specific antibodies, we show that the protein accumulates in rice leaves inoculated with P. syringae pv. syringae. Both the recombinant and the authentic proteins have an apparent molecular mass of 32 kDa (28.8 kDa calculated) and seem to be active as monomers. Pir7b esterase also exhibits sequence similarity to several expressed sequence tags of Arabidopsis thaliana, indicating that it belongs to a family of proteins widely occuring in plants.
Wäspi
U
U
Institut für Planzenbiologie, Universität Zürich, Switzerland.
Misteli
B
B
Hasslacher
M
M
Jandrositz
A
A
Kohlwein
S D
SD
Schwab
H
H
Dudler
R
R
D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D004950:Esterases / Q000737:chemistry*; D004952:Esters / Q000378:metabolism; D005658:Fungi / Q000472:pathogenicity; D008969:Molecular Sequence Data; D015394:Molecular Structure; D009284:Naphthols / Q000378:metabolism; D012275:Oryza / Q000201:enzymology / Q000469:parasitology*; D010935:Plant Diseases / Q000382:microbiology*; D010940:Plant Proteins / Q000737:chemistry*; D011549:Pseudomonas / Q000472:pathogenicity; D011994:Recombinant Proteins; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
D004952:Esters; D009284:Naphthols; D010940:Plant Proteins; D011994:Recombinant Proteins; D004950:Esterases; C113255:Pir7B protein, Oryza sativa; C008584:naphthol AS