The octadecanoid pathway is required for pathogen-induced multi-functional acetyl-CoA carboxylase accumulation in common bean (Phaseolus vulgaris L.).

A partial cDNA clone corresponding to the multi-functional acetyl-CoA carboxylase (ACCase, EC 6.4.1.2) was isolated using RNA extracted from methyl jasmonate (MeJA)-induced common bean cell cultures. Most of this clone corresponds to the 3' untranslated region and it showed high identity to alfalfa and soybean ACCase sequences. Southern hybridization revealed one copy of this gene in the common bean genome. In addition to being induced by MeJA in cell cultures and leaves, ACCase mRNA accumulated after yeast elicitor or Pseudomonas syringae pv tabaci treatment. Inhibitors of the octadecanoid pathway severely reduced ACCase mRNA and protein accumulation induced by yeast elicitor or P. syringae pv tabaci, indicating that jasmonates or a precursor mediate ACCase induction after pathogen infection. These results provide a role for the eukaryotic ACCase during the defense response to pathogens in common bean.

On the interaction of mitochondrial complex III with the Rieske iron-sulfur protein (subunit V).

Limited proteolysis of solubilized beef heart mitochondrial complex III with trypsin yields a product previously identified as fragment V" (González-Halphen, D., Lindorfer, M. A., and Capaldi, R. A. (1988) Biochemistry 27, 7021-7031). In this work, fragment V" was generated by trypsin treatment of both the intact complex III and the purified Rieske iron-sulfur protein. Thus, in its bound or isolated form, the same sites of subunit V are sensitive to protease action. Fragment V" was a soluble protein that retained its iron-sulfur moiety. It was purified by exclusion from a hydrophobic phenyl-Sepharose CL-4B column followed by gel filtration. In contrast to the pure, intact subunit V, fragment V" did not reconstitute oxidoreductase activity when combined with complex III devoid of subunit V. However, a 20-amino acid synthetic peptide carrying the sequence between amino acids Lys33 and Lys52 of the Rieske iron-sulfur protein competed with intact subunit V in reconstitution assays. The results obtained suggest that the iron-sulfur protein binds to complex III by hydrophobic protein-protein interactions, and that a nontransmembrane 18-amino acid amphipathic stretch accounts for the association of this subunit to the rest of the complex.