The tobacco salicylic acid-binding protein 3 (SABP3) is the chloroplast carbonic anhydrase, which exhibits antioxidant activity and plays a role in the hypersensitive defense response.
In plants, salicylic acid (SA) plays an important role in signaling both local and systemic defense responses. Previous efforts to identify SA effector proteins in tobacco have led to the isolation of two soluble cytoplasmic SA-binding proteins (SABPs): catalase, SABP, and an approximately 25-kDa protein, SABP2. Here we describe the identification of an SA-binding protein, SABP3, in the stroma of tobacco chloroplasts. SABP3 bound SA with an apparent dissociation constant (K(d)) of 3.7 microM and exhibited much greater affinity for biologically active than inactive analogs. Purification and partial sequencing of SABP3 indicated that it is the chloroplast carbonic anhydrase (CA). Confirming this finding, recombinant tobacco chloroplast CA exhibited both CA enzymatic and SA-binding activities. Expression of this protein in yeast also demonstrated that CA/SABP3 has antioxidant activity. A second gene encoding CA was also cloned, and its encoded protein was shown to behave similarly to that purified as SABP3. Finally, silencing of CA gene expression in leaves suppressed the Pto:avrPto-mediated hypersensitive response in disease resistance. These results demonstrate that SA may act through multiple effector proteins in plants and shed further light on the function of CA in chloroplasts.
Waksman Institute, Rutgers, State University of New Jersey, 190 Frelinghuysen Road, Piscataway, NJ 08854, USA.
David H
DH
Slaymaker
Duroy A
DA
Navarre
Daniel
D
Clark
Olga
O
del Pozo
Gregory B
GB
Martin
Daniel F
DF
Klessig
D000975:Antioxidants / Q000378:metabolism*; D001483:Base Sequence; D002256:Carbonic Anhydrases / Q000235:genetics / Q000378:metabolism*; D002352:Carrier Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D002736:Chloroplasts / Q000201:enzymology*; D003001:Cloning, Molecular; D017931:DNA Primers; D020868:Gene Silencing / Q000502:physiology; D005816:Genetic Complementation Test; D008969:Molecular Sequence Data; D010940:Plant Proteins; D017863:Potexvirus / Q000502:physiology; D014026:Nicotiana / Q000201:enzymology / Q000378:metabolism*
D000975:Antioxidants; D002352:Carrier Proteins; D017931:DNA Primers; D010940:Plant Proteins; D002256:Carbonic Anhydrases; C499505:SABP3 protein, Nicotiana tabacum