Bound nucleotide can control the dynamic architecture of monomeric actin

Polymerization of actin into cytoskeletal filaments is coupled to its bound adenine nucleotides. The mechanism by which nucleotide modulates actin functions has not been evident from analyses of ATP- and ADP-bound crystal structures of the actin monomer. We report that NMR chemical shift differences between the two forms are globally distributed. Furthermore, microsecond-millisecond motions are spread throughout the molecule in the ATP form, but largely confined to subdomains 1 and 2, and the nucleotide binding site in the ADP form. Through these motions, the ATP- and ADP-bound forms sample different high-energy conformations. A deafness-causing, fast-nucleating actin mutant populates the high-energy conformer of ATP-actin more than the wild-type protein, suggesting that this conformer may be on the pathway to nucleation. Together, the data suggest a model in which differential sampling of a nucleation-compatible form of the actin monomer may contribute to control of actin filament dynamics by nucleotide. NMR shows that ATP- and ADP-actin differ globally, including ground and excited state structures and dynamic architecture. Analyses of an actin mutant suggest the high-energy conformer of ATP-actin may be on the pathway to filament nucleation.

Automated summary

The polymerization of actin into cytoskeletal filaments is influenced by its bound adenine nucleotides. Through NMR analysis, it is found that ATP- and ADP-bound actin exhibit global differences in structure and dynamics. A deafness-causing actin mutant is found to predominantly populate the high-energy conformation of ATP-actin, suggesting its involvement in the nucleation of filaments. Collectively, these findings suggest that sampling of a nucleation-compatible form of the actin monomer may contribute to the regulation of actin filament dynamics by nucleotide.