Stability-activity tradeoffs constrain the adaptive evolution of RubisCO

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 111; no. 6; pp. 2223 - 2228
• Main Authors: Studer, Romain A., Christin, Pascal-Antoine, Williams, Mark A., Orengo, Christine A.
• Format: Journal Article
• Language: English
• Published: United States National Academy of Sciences 11.02.2014; National Acad Sciences
• Subjects: Active sites; Adaptation, Physiological; Amino acids; Angiospermae; Binding sites; Biological adaptation; Biological Evolution; Biological Sciences; Carbon Dioxide - metabolism; convergent evolution; Dimers; Ecological adaptation; Enzyme Stability; Enzymes; Evolution; evolutionary adaptation; Flowering plants; Flowers & plants; Genetic mutation; Models, Molecular; Molecular structure; Mutation; Photosynthesis; Phylogenetics; Phylogeny; Plant Physiological Phenomena; Plants; Positive selection; ribulose-bisphosphate carboxylase; Ribulose-Bisphosphate Carboxylase - chemistry; Ribulose-Bisphosphate Carboxylase - genetics; Ribulose-Bisphosphate Carboxylase - physiology
• DOI: 10.1073/pnas.1310811111
• ISSN: 0027-8424; 1091-6490

A well-known case of evolutionary adaptation is that of ribulose-1,5-bisphosphate carboxylase (RubisCO), the enzyme responsible for fixation of CO ₂ during photosynthesis. Although the majority of plants use the ancestral C ₃ photosynthetic pathway, many flowering plants have evolved a derived pathway named C ₄ photosynthesis. The latter concentrates CO ₂, and C ₄ RubisCOs consequently have lower specificity for, and faster turnover of, CO ₂. The C ₄ forms result from convergent evolution in multiple clades, with substitutions at a small number of sites under positive selection. To understand the physical constraints on these evolutionary changes, we reconstructed in silico ancestral sequences and 3D structures of RubisCO from a large group of related C ₃ and C ₄ species. We were able to precisely track their past evolutionary trajectories, identify mutations on each branch of the phylogeny, and evaluate their stability effect. We show that RubisCO evolution has been constrained by stability-activity tradeoffs similar in character to those previously identified in laboratory-based experiments. The C ₄ properties require a subset of several ancestral destabilizing mutations, which from their location in the structure are inferred to mainly be involved in enhancing conformational flexibility of the open-closed transition in the catalytic cycle. These mutations are near, but not in, the active site or at intersubunit interfaces. The C ₃ to C ₄ transition is preceded by a sustained period in which stability of the enzyme is increased, creating the capacity to accept the functionally necessary destabilizing mutations, and is immediately followed by compensatory mutations that restore global stability.