Hsc70 is required for endocytosis and clathrin function in Drosophila

Henry C. Chang, Sherri L. Newmyer, Michael J. Hull, Melanie Ebersold, Sandra L. Schmid, Ira Mellman

Research output: Contribution to journalArticle

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Abstract

By screening for Drosophila mutants exhibiting aberrant bride of sevenless (Boss) staining patterns on eye imaginal disc epithelia, we have recovered a point mutation in Hsc70-4, the closest homologue to bovine clathrin uncoating ATPase. Although the mutant allele was lethal, analysis of mutant clones generated by FLP/FRT recombination demonstrated that the Sevenless-mediated internalization of Boss was blocked in mutant Hsc70-4 eye disc epithelial cells. Endocytosis of other probes was also greatly inhibited in larval Garland cells. Immunostaining and EM analysis of the mutant cells revealed disruptions in the organization of endosomal/lysosomal compartments, including a substantial reduction in the number of clathrin-coated structures in Garland cells. The Hsc70-4 mutation also interacted genetically with a dominant-negative mutant of dynamin, a gene required for the budding of clathrin-coated vesicles (CCVs). Consistent with these phenotypes, recombinant mutant Hsc70 proteins exhibited diminished clathrin uncoating activity in vitro. Together, these data provide genetic support for the long-suspected role of Hsc70 in clathrin-mediated endocytosis, at least in part by inhibiting the uncoating of CCVs.

Original languageEnglish (US)
Pages (from-to)477-487
Number of pages11
JournalJournal of Cell Biology
Volume159
Issue number3
DOIs
StatePublished - Nov 11 2002

Fingerprint

Clathrin
Endocytosis
Drosophila
Clathrin-Coated Vesicles
HSC70 Heat-Shock Proteins
Lethal Genes
Imaginal Discs
Dynamins
Mutant Proteins
Point Mutation
Recombinant Proteins
Genetic Recombination
Adenosine Triphosphatases
Epithelium
Clone Cells
Epithelial Cells
Staining and Labeling
Phenotype
Mutation
Genes

Keywords

  • Boss internalization
  • Clathrin uncoating
  • Drosophila
  • Endocytosis
  • Hsc70

ASJC Scopus subject areas

  • Cell Biology

Cite this

Chang, H. C., Newmyer, S. L., Hull, M. J., Ebersold, M., Schmid, S. L., & Mellman, I. (2002). Hsc70 is required for endocytosis and clathrin function in Drosophila. Journal of Cell Biology, 159(3), 477-487. https://doi.org/10.1083/jcb.200205086

Hsc70 is required for endocytosis and clathrin function in Drosophila. / Chang, Henry C.; Newmyer, Sherri L.; Hull, Michael J.; Ebersold, Melanie; Schmid, Sandra L.; Mellman, Ira.

In: Journal of Cell Biology, Vol. 159, No. 3, 11.11.2002, p. 477-487.

Research output: Contribution to journalArticle

Chang, HC, Newmyer, SL, Hull, MJ, Ebersold, M, Schmid, SL & Mellman, I 2002, 'Hsc70 is required for endocytosis and clathrin function in Drosophila', Journal of Cell Biology, vol. 159, no. 3, pp. 477-487. https://doi.org/10.1083/jcb.200205086
Chang, Henry C. ; Newmyer, Sherri L. ; Hull, Michael J. ; Ebersold, Melanie ; Schmid, Sandra L. ; Mellman, Ira. / Hsc70 is required for endocytosis and clathrin function in Drosophila. In: Journal of Cell Biology. 2002 ; Vol. 159, No. 3. pp. 477-487.
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