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PFN2 and NAA80 cooperate to efficiently acetylate the N-terminus of actin.
- Source :
-
The Journal of biological chemistry [J Biol Chem] 2020 Dec 04; Vol. 295 (49), pp. 16713-16731. Date of Electronic Publication: 2020 Sep 25. - Publication Year :
- 2020
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Abstract
- 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.<br />Competing Interests: Conflict of interest—The authors declare that they have no conflicts of interest with the contents of this article.<br /> (© 2020 Ree et al.)
- Subjects :
- Acetylation
Acetyltransferases chemistry
Acetyltransferases genetics
Actin Cytoskeleton metabolism
Actins chemistry
Animals
Biocatalysis
Cell Line
Humans
Profilins chemistry
Profilins deficiency
Profilins genetics
Protein Binding
Protein Domains
Protein Structure, Quaternary
Recombinant Proteins biosynthesis
Recombinant Proteins chemistry
Recombinant Proteins isolation & purification
Scattering, Small Angle
Ultracentrifugation
X-Ray Diffraction
Acetyltransferases metabolism
Actins metabolism
Profilins metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1083-351X
- Volume :
- 295
- Issue :
- 49
- Database :
- MEDLINE
- Journal :
- The Journal of biological chemistry
- Publication Type :
- Academic Journal
- Accession number :
- 32978259
- Full Text :
- https://doi.org/10.1074/jbc.RA120.015468