The Journal of Cell Biology, Vol 112, 65-80, Copyright © 1991 by The Rockefeller University Press

ARTICLES

Sequence of the clathrin heavy chain from Saccharomyces cerevisiae and requirement of the COOH terminus for clathrin function

SK Lemmon, A Pellicena-Palle, K Conley and CL Freund
Department of Molecular Biology and Microbiology, Case Western Reserve University School of Medicine, Cleveland, Ohio 44106.

The sequence of the clathrin heavy chain gene, CHC1, from Saccharomyces cerevisiae is reported. The gene encodes a protein of 1,653 amino acids that is 50% identical to the rat clathrin heavy chain (HC) (Kirchhausen, T., S. C. Harrison, E. P. Chow, R. J. Mattaliano, R. L. Ramachandran, J. Smart, and J. Brosius. 1987. Proc. Natl. Acad. Sci. USA. 84:8805-8809). The alignment extends over the complete length of the two proteins, except for a COOH-terminal extension of the rat HC and a few small gaps, primarily in the globular terminal domain. The yeast HC has four prolines in the region of the rat polypeptide that was proposed to form the binding site for clathrin light chains via an alpha-helical coiled-coil interaction. The yeast protein also lacks the COOH-terminal Pro-Gly rich segment present in the last 45 residues of the rat HC, which were proposed to be involved in the noncovalent association of HCs to form trimers at the triskelion vertex. To examine the importance of the COOH terminus of the HC for clathrin function, a HC containing a COOH-terminal deletion of 57 amino acids (HC delta 57) was expressed in clathrin-deficient yeast (chc1-delta). HC delta 57 rescued some of the phenotypes (slow growth at 30 degrees, genetic instability, and defects in mating and sporulation) associated with the chc1-delta mutation to normal or near normal. Also, truncated HCs were assembled into triskelions. However, cells with HC delta 57 were temperature sensitive for growth and still displayed a major defect in processing of the mating pheromone alpha-factor. Fewer coated vesicles could be isolated from cells with HC delta 57 than cells with the wild- type HC. This suggests that the COOH-terminal region is not required for formation of trimers, but it may be important for normal clathrin- coated vesicle structure and function.
CiteULike    Complore    Connotea    Del.icio.us    Digg    Facebook    Reddit    Technorati    Twitter    What's this?

This article has been cited by other articles:

• Collette, J. R., Chi, R. J., Boettner, D. R., Fernandez-Golbano, I. M., Plemel, R., Merz, A. J., Geli, M. I., Traub, L. M., Lemmon, S. K. (2009). Clathrin Functions in the Absence of the Terminal Domain Binding Site for Adaptor-associated Clathrin-Box Motifs. Mol. Biol. Cell 20: 3401-3413 [Abstract] [Full Text]  
• Sato, K., Ernstrom, G. G., Watanabe, S., Weimer, R. M., Chen, C.-H., Sato, M., Siddiqui, A., Jorgensen, E. M., Grant, B. D. (2009). Differential requirements for clathrin in receptor-mediated endocytosis and maintenance of synaptic vesicle pools. Proc. Natl. Acad. Sci. USA 106: 1139-1144 [Abstract] [Full Text]  
• Bensen, E. S., Costaguta, G., Payne, G. S. (2000). Synthetic Genetic Interactions With Temperature-Sensitive Clathrin in Saccharomyces cerevisiae: Roles for Synaptojanin-Like Inp53p and Dynamin-Related Vps1p in Clathrin-Dependent Protein Sorting at the trans-Golgi Network. Genetics 154: 83-97 [Abstract] [Full Text]  
• Chu, D. S., Pishvaee, B., Payne, G. S. (1999). A Modulatory Role for Clathrin Light Chain Phosphorylation in Golgi Membrane Protein Localization during Vegetative Growth and during the Mating Response of Saccharomyces cerevisiae. Mol. Biol. Cell 10: 713-726 [Abstract] [Full Text]  
• Chen, C.-Y., Graham, T. R. (1998). An arf1{Delta} Synthetic Lethal Screen Identifies a New Clathrin Heavy Chain Conditional Allele That Perturbs Vacuolar Protein Transport in Saccharomyces cerevisiae. Genetics 150: 577-589 [Abstract] [Full Text]  
• Liu, S.-H., Marks, M. S., Brodsky, F. M. (1998). A Dominant-negative Clathrin Mutant Differentially Affects Trafficking of Molecules with Distinct Sorting Motifs in the Class II Major Histocompatibility Complex (MHC) Pathway. JCB 140: 1023-1037 [Abstract] [Full Text]  
• Kokoza, V. A., Raikhel, A. S. (1997). Ovarian- and Somatic-specific Transcripts of the Mosquito Clathrin Heavy Chain Gene Generated by Alternative 5'-Exon Splicing and Polyadenylation. J. Biol. Chem. 272: 1164-1170 [Abstract] [Full Text]  
• Huang, K., Gullberg, L, Nelson, K., Stefan, C., Blumer, K, Lemmon, S. (1997). Novel functions of clathrin light chains: clathrin heavy chain trimerization is defective in light chain-deficient yeast. J. Cell Sci. 110: 899-910 [Abstract]  
• Blackbourn, H., Jackson, A. (1996). Plant clathrin heavy chain: sequence analysis and restricted localisation in growing pollen tubes. J. Cell Sci. 109: 777-786 [Abstract]  

Home | Help | Feedback | Subscriptions | Archive | Search | Table of Contents