Back to Search
Start Over
Clathrin Functions in the Absence of the Terminal Domain Binding Site for Adaptor-associated Clathrin-Box Motifs
- Source :
- Molecular Biology of the Cell. 20:3401-3413
- Publication Year :
- 2009
- Publisher :
- American Society for Cell Biology (ASCB), 2009.
-
Abstract
- Clathrin is involved in vesicle formation in the trans-Golgi network (TGN)/endosomal system and during endocytosis. Clathrin recruitment to membranes is mediated by the clathrin heavy chain (HC) N-terminal domain (TD), which forms a seven-bladed β-propeller. TD binds membrane-associated adaptors, which have short peptide motifs, either the clathrin-box (CBM) and/or the W-box; however, the importance of the TD binding sites for these motifs has not been tested in vivo. We investigated the importance of the TD in clathrin function by generating 1) mutations in the yeast HC gene (CHC1) to disrupt the binding sites for the CBM and W-box (chc1-box), and 2) four TD-specific temperature-sensitive alleles of CHC1. We found that TD is important for the retention of resident TGN enzymes and endocytosis of α-factor; however, the known adaptor binding sites are not necessary, because chc1-box caused little to no effect on trafficking pathways involving clathrin. The Chc1-box TD was able to interact with the endocytic adaptor Ent2 in a CBM-dependent manner, and HCs encoded by chc1-box formed clathrin-coated vesicles. These data suggest that additional or alternative binding sites exist on the TD propeller to help facilitate the recruitment of clathrin to sites of vesicle formation.
- Subjects :
- Saccharomyces cerevisiae Proteins
Endosome
Recombinant Fusion Proteins
Amino Acid Motifs
Green Fluorescent Proteins
Molecular Sequence Data
Endocytic cycle
Saccharomyces cerevisiae
Plasma protein binding
Endocytosis
Aminopeptidases
Clathrin
Protein Structure, Secondary
Clathrin Heavy Chains
Amino Acid Sequence
Molecular Biology
Alleles
Chitin Synthase
Binding Sites
biology
Vesicle
Temperature
Clathrin-Coated Vesicles
Articles
Cell Biology
Cell biology
Adaptor Proteins, Vesicular Transport
Protein Transport
Biochemistry
Mutation
biology.protein
Clathrin adaptor proteins
Protein Processing, Post-Translational
Protein Binding
trans-Golgi Network
Subjects
Details
- ISSN :
- 19394586 and 10591524
- Volume :
- 20
- Database :
- OpenAIRE
- Journal :
- Molecular Biology of the Cell
- Accession number :
- edsair.doi.dedup.....eed3aa7a1c1c811bd2d0e2b382a5afcf