Multivesicular Body Sorting: Ubiquitin Ligase Rsp5 Is Required for the Modification and Sorting of Carboxypeptidase S

David J Katzmann, Srimonti Sarkar, Tony Chu, Anjon Audhya, Scott D. Emr

Research output: Contribution to journalArticle

117 Citations (Scopus)

Abstract

The multivesicular body (MVB) sorting pathway provides a mechanism for delivering transmembrane proteins into the lumen of the lysosome/vacuole. Recent studies demonstrated that ubiquitin modification acts in cis as a signal for the sorting of cargoes into this pathway. Here, we present results from a genetic selection designed to identify mutants that missort MVB cargoes. This selection identified a point mutation in ubiquitin ligase Rsp5 (Rsp5-326). At the permissive temperature, this mutant is specifically defective for ubiquitination and sorting of the ubiquitin-dependent MVB cargo precursor carboxypeptidase S (pCPS), but not ligand-induced ubiquitination of Ste2. A previous study implicated Tul1 as the ubiquitin ligase responsible for MVB sorting of pCPS. However, we detected no defect in either the sorting or ubiquitination of pCPS in tul1 mutants. We had previously shown that Fab1 phosphatidylinositol 3-phosphate 5-kinase is also required for MVB sorting of pCPS, but not Ste2. However, our analyses reveal that fab1 mutants do not exhibit a defect in ubiquitination of pCPS. Thus, both Rsp5 and Fab1 play distinct and essential roles in the targeting of biosynthetic MVB cargoes. However, whereas Rsp5 seems to be responsible for cargo ubiquitination, the precise role for Fab1 remains to be elucidated.

Original languageEnglish (US)
Pages (from-to)468-480
Number of pages13
JournalMolecular Biology of the Cell
Volume15
Issue number2
DOIs
StatePublished - Feb 2004

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Multivesicular Bodies
Carboxypeptidases
Ligases
Ubiquitin
Ubiquitination
Genetic Selection
Vacuoles
Lysosomes
Point Mutation
Phosphotransferases
Ligands
Temperature
Proteins

ASJC Scopus subject areas

  • Cell Biology
  • Genetics
  • Molecular Biology

Cite this

Multivesicular Body Sorting : Ubiquitin Ligase Rsp5 Is Required for the Modification and Sorting of Carboxypeptidase S. / Katzmann, David J; Sarkar, Srimonti; Chu, Tony; Audhya, Anjon; Emr, Scott D.

In: Molecular Biology of the Cell, Vol. 15, No. 2, 02.2004, p. 468-480.

Research output: Contribution to journalArticle

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