PFN2 and NAA80 cooperate to efficiently acetylate the N-terminus of actin
| Autor: | Ree, Rasmus, Kind, Laura, Kaziales, Anna, Varland, Sylvia, Dai, Minglu, Richter, Klaus, Drazic, Adrian, Arnesen, Thomas |
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| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
post-translational modification (PTM) small-angle X-ray scattering (SAXS) Biochemistry protein-protein interaction Profilins X-Ray Diffraction Cytoskeleton 0303 health sciences biology Chemistry 030302 biochemistry & molecular biology cytoskeleton Recombinant Proteins Cell biology Actin Cytoskeleton Profilin Acetyltransferase Protein Structure and Folding actin Intracellular Protein Binding macromolecular substances Cell Line Protein–protein interaction 03 medical and health sciences Protein Domains Acetyltransferases Profilin-1 Scattering Small Angle Animals Humans profilin Protein Structure Quaternary Molecular Biology Actin acetylation 030304 developmental biology 030102 biochemistry & molecular biology N-terminal acetyltransferases Cell Biology Actin cytoskeleton Actins N-terminus 030104 developmental biology Acetylation Biocatalysis Enzymology biology.protein NAT Ultracentrifugation |
| Zdroj: | Journal of Biological Chemistry 16713-16731 The Journal of Biological Chemistry |
| ISSN: | 0021-9258 |
| Popis: | The actin cytoskeleton is of profound importance to cell shape, division, and intracellular force generation. Profilins bind to globular (G-)actin and regulate actin filament formation. Although profilins are well-established actin regulators, the distinct roles of the dominant profilin, profilin 1 (PFN1), versus the less abundant profilin 2 (PFN2) remain enigmatic. In this study, we use interaction proteomics to discover that PFN2 is an interaction partner of the actin N-terminal acetyltransferase NAA80, and further confirm this by analytical ultracentrifugation. Enzyme assays with NAA80 and different profilins demonstrate that PFN2 binding specifically increases the intrinsic catalytic activity of NAA80. NAA80 binds PFN2 through a proline-rich loop, deletion of which abrogates PFN2 binding. Small-angle X-ray scattering shows that NAA80, actin, and PFN2 form a ternary complex and that NAA80 has partly disordered regions in the N-terminus and the proline-rich loop, the latter of which is partly ordered upon PFN2 binding. Furthermore, binding of PFN2 to NAA80 via the proline-rich loop promotes binding between the globular domains of actin and NAA80, and thus acetylation of actin. However, the majority of cellular NAA80 is stably bound to PFN2 and not to actin, and we propose that this complex acetylates G-actin before it is incorporated into filaments. In conclusion, we reveal a functionally specific role of PFN2 as a stable interactor and regulator of the actin N-terminal acetyltransferase NAA80, and establish the modus operandi for NAA80-mediated actin N-terminal acetylation, a modification with a major impact on cytoskeletal dynamics. publishedVersion |
| Databáze: | OpenAIRE |
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