Heterodimeric interactions among the 1-amino-cyclopropane-1-carboxylate synthase polypeptides encoded by the Arabidopsis gene family.
The pyridoxal phosphate-dependent enzyme, 1-aminocyclopropane-1-carboxylate synthase (ACS; EC 4.4.1.14), catalyzes the rate-limiting step in the ethylene biosynthetic pathway in plants. The Arabidopsis genome encodes nine ACS polypeptides that form eight functional (ACS2, ACS4-9, ACS11) and one nonfunctional (ACS1) homodimers. Because the enzyme is a homodimer with shared active sites, the question arises whether the various polypeptides can form functional heterodimers. Intermolecular complementation experiments in Escherichia coli by coexpressing the K278A and Y92A mutants of different polypeptides show that all of them have the capacity to heterodimerize. However, functional heterodimers are formed only among gene family members that belong to one or the other of the two phylogenetic branches. ACS7 is an exception to this rule, which forms functional heterodimers with some members of both branches when it provides the wt K278 residue. ACS1, the nonfunctional polypeptide as a homodimer, can also form functional heterodimers with members of its phylogenetic branch when its partners provide the wt K278 residue. The ACS gene family products can potentially form 45 homo- and heterodimers of which 25 are functional. Bimolecular fluorescence complementation and biochemical coaffinity purification assays show that the inactivity of certain heterodimers is not due to the absence of heterodimerization but rather to structural restraint(s) that prevents the shared active sites from being functional. We propose that functional heterodimerization enhances the isozyme diversity of the ACS gene family and provides physiological versatility by being able to operate in a broad gradient of S-adenosylmethionine concentration in various cells/tissues during plant growth and development. Nonfunctional heterodimerization may also play a regulatory role during the plant life cycle.
Tsuchisaka
Atsunari
A
Plant Gene Expression Center, 800 Buchanan Street, Albany, CA 94710, USA.
Theologis
Athanasios
A
D017360:Arabidopsis / Q000201:enzymology / Q000235:genetics*; D029681:Arabidopsis Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D003001:Cloning, Molecular; D019281:Dimerization; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics / Q000254:growth & development; D005816:Genetic Complementation Test; D008190:Lyases / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D005810:Multigene Family; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism
D029681:Arabidopsis Proteins; D011994:Recombinant Proteins; D008190:Lyases; C021773:1-aminocyclopropanecarboxylate synthase