The ubiquitin E3 ligase POSH regulates calcium homeostasis through spatial control of Herp.

The Journal of cell biology ·Vol. 177 ·No. 1 ·2007-04-09 ·Pages 51-61

Tuvia S, Taglicht D, Erez O, Alroy I, Alchanati I, Bicoviski V, Dori-Bachash M, Ben-Avraham D, Reiss Y

Abstract

The ubiquitin (Ub) domain protein Herp plays a crucial role in the maintenance of calcium homeostasis during endoplasmic reticulum (ER) stress. We now show that Herp is a substrate as well as an activator of the E3 Ub ligase POSH. Herp-mediated POSH activation requires the Ubl domain and exclusively promotes lysine-63-linked polyubiquitination. Confocal microscopy demonstrates that Herp resides mostly in the trans-Golgi network, but, shortly after calcium perturbation by thapsigargin (Tpg), it appears mainly in the ER. Substitution of all lysine residues within the Ubl domain abolishes lysine-63-linked polyubiquitination of Herp in vitro and calcium-induced Herp relocalization that is also abrogated by the overexpression of a dominant-negative POSHV14A. A correlation exists between the kinetics of Tpg-induced Herp relocalization and POSH-dependent polyubiquitination. Finally, the overexpression of POSH attenuates, whereas the inhibition of POSH by the expression of POSHV14A or by RNA interference enhances Tpg-induced calcium burst. Altogether, these results establish a critical role for POSH-mediated ubiquitination in the maintenance of calcium homeostasis through the spatial control of Herp.

MeSH Terms

Calcium/metabolism Endoplasmic Reticulum/metabolism Enzyme Inhibitors/pharmacology HeLa Cells Homeostasis Humans Intracellular Membranes/metabolism Membrane Proteins/chemistry,metabolism Models, Biological Protein Structure, Tertiary Thapsigargin/pharmacology Tunicamycin/pharmacology Ubiquitin/metabolism Ubiquitin-Protein Ligases/metabolism,physiology trans-Golgi Network/metabolism

Chemicals

Enzyme Inhibitors HERPUD1 protein, human Membrane Proteins Ubiquitin Tunicamycin Thapsigargin SH3RF1 protein, human Ubiquitin-Protein Ligases Calcium

Authors & Affiliations

9 authors, click to expand affiliations / ORCID

Tuvia Shmuel

Proteologics Ltd., Kiryat Weizmann, Rehovot 76124, Israel.

Taglicht Daniel

Erez Omri

Alroy Iris

Alchanati Iris

Bicoviski Vivian

Dori-Bachash Mally

Ben-Avraham Danny

Reiss Yuval

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Article Info

Journal

The Journal of cell biology

Abbr.

J Cell Biol

ISSN

0021-9525

DOI

10.1083/jcb.200611036

Published

2007-04-09

Pages

51-61

Language

English

Region

United States

NLM ID

0375356

PMCID

PMC2064109

Subset

External Links

PubMed source DOI: 10.1083/jcb.200611036 PMC Full Text View journal details

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The ubiquitin E3 ligase POSH regulates calcium homeostasis through spatial control of Herp.

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