Vps41p function in the alkaline phosphatase pathway requires homo-oligomerization and interaction with AP-3 through two distinct domains

T. Darsow, David J Katzmann, C. R. Cowles, S. D. Emr

Research output: Contribution to journalArticle

61 Citations (Scopus)

Abstract

Transport of proteins through the ALP (alkaline phosphatase) pathway to the vacuole requires the function of the AP-3 adaptor complex and Vps41p. However, unlike other adaptor protein-dependent pathways, the ALP pathway has not been shown to require additional accessory proteins or coat proteins, such as membrane recruitment factors or clathrin. Two independent genetic approaches have been used to identify new mutants that affect transport through the ALP pathway. These screens yielded new mutants in both VPS41 and the four AP-3 subunit genes. Two new VPS41 alleles exhibited phenotypes distinct from null mutants of VPS41, which are defective in vacuolar morphology and protein transport through both the ALP and CPY sorting pathways. The new alleles displayed severe ALP sorting defects, normal vacuolar morphology, and defects in ALP vesicle formation at the Golgi complex. Sequencing analysis of these VPS41 alleles revealed mutations encoding amino acid changes in two distinct domains of Vps41p: a conserved N-terminal domain and a C-terminal clathrin heavy-chain repeat (CHCR) domain. We demonstrate that the N-terminus of Vps41p is required for binding to AP-3, whereas the C-terminal CHCR domain directs homo-oligomerization of Vps41p. These data indicate that a homo-oligomeric form of Vps41p is required for the formation of ALP containing vesicles at the Golgi complex via interactions with AP-3.

Original languageEnglish (US)
Pages (from-to)37-51
Number of pages15
JournalMolecular Biology of the Cell
Volume12
Issue number1
StatePublished - 2001
Externally publishedYes

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Alkaline Phosphatase
Clathrin Heavy Chains
Alleles
Golgi Apparatus
Clathrin
Nucleic Acid Repetitive Sequences
Capsid Proteins
Protein Transport
Vacuoles
Carrier Proteins
Proteins
Phenotype
Amino Acids
Mutation
Membranes
Genes

ASJC Scopus subject areas

  • Cell Biology
  • Genetics
  • Molecular Biology

Cite this

Vps41p function in the alkaline phosphatase pathway requires homo-oligomerization and interaction with AP-3 through two distinct domains. / Darsow, T.; Katzmann, David J; Cowles, C. R.; Emr, S. D.

In: Molecular Biology of the Cell, Vol. 12, No. 1, 2001, p. 37-51.

Research output: Contribution to journalArticle

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AU - Emr, S. D.

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