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PMID: 10637223 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Monoubiquitin carries a novel internalization signal that is appended to activated receptors.

The EMBO journal ·Vol. 19 ·No. 2 ·2000-01-17 ·Pages 187-98

Shih SC, Sloper-Mould KE, Hicke L

Abstract

Ubiquitin modification of signal transducing receptors at the plasma membrane is necessary for rapid receptor internalization and downregulation. We have investigated whether ubiquitylation alters a receptor cytoplasmic tail to reveal a previously masked internalization signal, or whether ubiquitin itself carries an internalization signal. Using an alpha-factor receptor-ubiquitin chimeric protein, we demonstrate that monoubiquitin can mediate internalization of an activated receptor that lacks all cytoplasmic tail sequences. Furthermore, fusion of ubiquitin in-frame to the stable plasma membrane protein Pma1p stimulates endocytosis of this protein. Ubiquitin does not carry a functional tyrosine- or di-leucine-based internalization signal. Instead, the three-dimensional structure of the folded ubiquitin polypeptide carries an internalization signal that consists of two surface patches surrounding the critical residues Phe4 and Ile44. We conclude that ubiquitin functions as a novel regulated internalization signal that can be appended to a plasma membrane protein to trigger downregulation.

MeSH Terms
Amino Acid Sequence Amino Acid Substitution Cell Membrane/metabolism Isoenzymes/chemistry,metabolism Isoleucine Kinetics Models, Molecular Molecular Sequence Data Mutagenesis, Site-Directed Phenylalanine Protein Conformation Proton-Translocating ATPases/chemistry,metabolism Receptors, Mating Factor Receptors, Peptide/chemistry,metabolism Recombinant Fusion Proteins/chemistry,metabolism Saccharomyces cerevisiae/genetics,metabolism Saccharomyces cerevisiae Proteins Sequence Deletion Signal Transduction Transcription Factors Ubiquitins/chemistry,metabolism
Chemicals
Isoenzymes PMA2 protein, S cerevisiae Receptors, Mating Factor Receptors, Peptide Recombinant Fusion Proteins Saccharomyces cerevisiae Proteins Transcription Factors Ubiquitins Isoleucine Phenylalanine PMA1 protein, S cerevisiae Proton-Translocating ATPases
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Shih S C
Department of Biochemistry, Molecular Biology and Cell Biology, Northwestern University, Evanston, IL 60208, USA.
Sloper-Mould K E
Hicke L
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1093/emboj/19.2.187
Published
2000-01-17
Pages
187-98
Language
English
Region
England
NLM ID
8208664
PMCID
PMC305553
Subset
IM
Grants
NIDDK NIH HHS · R01 DK053257 · United States
NIGMS NIH HHS · T32 GM008061 · United States
NIDDK NIH HHS · DK53257 · United States
NIGMS NIH HHS · T32GM08061 · United States
External Links
PubMed source DOI: 10.1093/emboj/19.2.187 PMC Full Text View journal details
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