title: A novel structural model for regulation of clathrin function.
abstract: The distinctive triskelion shape of clathrin allows assembly into polyhedral lattices during the process of clathrin-coated vesicle formation. We have used random and site-directed mutagenesis of the yeast clathrin heavy chain gene (CHC1) to characterize regions which determine Chc trimerization and binding to the clathrin light chain (Clc) subunit. Analysis of the mutants indicates that mutations in the trimerization domain at the triskelion vertex, as well as mutations in the adjacent leg domain, frequently influence Clc binding. Strikingly, one mutation in the trimerization domain enhances the association of Clc with Chc. Additional mutations in the trimerization domain, in combination with mutations in the adjacent leg domain, exhibit severe defects in Clc binding while maintaining near normal trimerization properties. The position of these trimerization domain mutations on one face of a putative alpha-helix defines a region on the trimer surface that interacts directly with Clc. These results suggest that Clc extends into the Chc trimerization domain from the adjacent leg, thereby bridging the two domains. On the basis of this conclusion, we propose a new model for the organization of the triskelion vertex which provides a structural basis for regulatory effects of Clc on clathrin function.
authors.affiliation: Department of Biological Chemistry, UCLA School of Medicine, Los Angeles, CA 90095, USA.
authors.forename: B
authors.initials: B
authors.lastname: Pishvaee
authors.forename: A
authors.initials: A
authors.lastname: Munn
authors.forename: G S
authors.initials: GS
authors.lastname: Payne
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002966:Clathrin / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D033922:Clathrin Heavy Chains; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D011487:Protein Conformation; D017510:Protein Folding; D051381:Rats; D012441:Saccharomyces cerevisiae; D013329:Structure-Activity Relationship; D013696:Temperature
chemical_list: D002966:Clathrin; D033922:Clathrin Heavy Chains