title: Nanostructured self-assembly of inverted formin 2 (INF2) and F-actin-INF2 complexes revealed by atomic force microscopy.
abstract: Self-organization of cytoskeletal proteins such as actin and tubulin into filaments and microtubules is frequently assisted by the proteins binding to them. Formins are regulatory proteins that nucleate the formation of new filaments and are essential for a wide range of cellular functions. The vertebrate inverted formin 2 (INF2) has both actin filament nucleating and severing/depolymerizing activities connected to its ability to encircle actin filaments. Using atomic force microscopy, we report that a formin homology 2 (FH2) domain-containing construct of INF2 (INF2-FH1-FH2-C or INF2-FFC) self-assembles into nanoscale ringlike oligomeric structures in the absence of actin filaments, demonstrating an inherent ability to reorganize from a dimeric to an oligomeric state. A construct lacking the C-terminal region (INF2-FH1-FH2 or INF2-FF) also oligomerizes, confirming the dominant role of FH2-mediated interactions. Moreover, INF2-FFC domains were observed to organize into ringlike structures around single actin filaments. This is the first demonstration that formin FH2 domains can self-assemble into oligomers in the absence of filaments and has important implications for observing unaveraged decoration and/or remodeling of filaments by actin binding proteins.
authors.affiliation: Department of Chemistry and Biochemistry, University of California , Los Angeles, California 90095, United States.
authors.forename: Shivani
authors.initials: S
authors.lastname: Sharma
authors.forename: Elena E
authors.initials: EE
authors.lastname: Grintsevich
authors.forename: JungReem
authors.initials: J
authors.lastname: Woo
authors.forename: Pinar S
authors.initials: PS
authors.lastname: Gurel
authors.forename: Henry N
authors.initials: HN
authors.lastname: Higgs
authors.forename: Emil
authors.initials: E
authors.lastname: Reisler
authors.forename: James K
authors.initials: JK
authors.lastname: Gimzewski
mesh_terms: D000199:Actins / Q000737:chemistry* / Q000378:metabolism*; D008840:Microfilament Proteins / Q000737:chemistry* / Q000378:metabolism*; D018625:Microscopy, Atomic Force / Q000379:methods*; D011485:Protein Binding
chemical_list: D000199:Actins; D008840:Microfilament Proteins