title: Interaction between microtubules and the Drosophila formin Cappuccino and its effect on actin assembly.
abstract: Formin family actin nucleators are potential coordinators of the actin and microtubule cytoskeletons, as they can both nucleate actin filaments and bind microtubules in vitro. To gain a more detailed mechanistic understanding of formin-microtubule interactions and formin-mediated actin-microtubule cross-talk, we studied microtubule binding by Cappuccino (Capu), a formin involved in regulating actin and microtubule organization during Drosophila oogenesis. We found that two distinct domains within Capu, FH2 and tail, work together to promote high-affinity microtubule binding. The tail domain appears to bind microtubules through nonspecific charge-based interactions. In contrast, distinct residues within the FH2 domain are important for microtubule binding. We also report the first visualization of a formin polymerizing actin filaments in the presence of microtubules. Interestingly, microtubules are potent inhibitors of the actin nucleation activity of Capu but appear to have little effect on Capu once it is bound to the barbed end of an elongating filament. Because Capu does not simultaneously bind microtubules and assemble actin filaments in vitro, its actin assembly and microtubule binding activities likely require spatial and/or temporal regulation within the Drosophila oocyte.
authors.affiliation: From the Molecular Biology Interdepartmental PhD Program.
authors.forename: Elizabeth A
authors.initials: EA
authors.lastname: Roth-Johnson
authors.forename: Christina L
authors.initials: CL
authors.lastname: Vizcarra
authors.forename: Justin S
authors.initials: JS
authors.lastname: Bois
authors.forename: Margot E
authors.initials: ME
authors.lastname: Quinlan
mesh_terms: D008841:Actin Cytoskeleton / Q000235:genetics / Q000378:metabolism; D000199:Actins / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D029721:Drosophila Proteins / Q000235:genetics / Q000378:metabolism*; D004331:Drosophila melanogaster; D005260:Female; D008297:Male; D008840:Microfilament Proteins / Q000235:genetics / Q000378:metabolism*; D008870:Microtubules / Q000235:genetics / Q000378:metabolism*; D009865:Oocytes / Q000166:cytology / Q000378:metabolism*; D009866:Oogenesis / Q000502:physiology*; D055503:Protein Multimerization / Q000502:physiology*; D017434:Protein Structure, Tertiary
chemical_list: D000199:Actins; D029721:Drosophila Proteins; D008840:Microfilament Proteins; C096354:capu protein, Drosophila
keywords: Actin; Capu; Cell Polarity; Cytoskeleton; Formin; Microtubules; Oocyte; TIRF