Tyrosine phosphorylation of p97 regulates transitional endoplasmic reticulum assembly in vitro

Lavoie, C., Chevet, E., Roy, L., Tonks, N. K., Fazel, A., Posner, B. I., Paiement, J., Bergeron, J. J. M. (December 2000) Tyrosine phosphorylation of p97 regulates transitional endoplasmic reticulum assembly in vitro. Proceedings of the National Academy of Sciences of the United States of America, 97 (25). pp. 13637-13642. ISSN 0027-8424

URL: http://www.ncbi.nlm.nih.gov/pubmed/11087817
DOI: 10.1073/pnas.240278097

Abstract

The ATPase associated with different cellular activities family member p97, associated p47, and the t-SNARE syntaxin 5 are necessary for the cell-free reconstitution of transitional endoplasmic reticulum (tER) from starting low-density microsomes. Here, we report that membrane-associated tyrosine kinase and protein-tyrosine phosphatase (PTPase) activities regulate tER assembly by stabilizing (PTPase) or destabilizing (tyrosine kinase) p97 association with membranes. Incubation with the PTPase inhibitor bpV-(phen) inhibited tER assembly coincident with the enhanced tyrosine phosphorylation of endogenous p97 and its release from membranes. By contrast, the tyrosine kinase inhibitor, genistein, promoted tER formation and prevented p97 dissociation from membranes while increasing p97 association with the t-SNARE syntaxin 5. Purification of the endogenous tyrosine kinase activity from low-density microsomes led to the identification of JAK-2, whereas PTPH1 was identified as the relevant PTPase. The p97 tyrosine phosphorylation state is proposed to coordinate the assembly of the tER as a regulatory step of the early secretory pathway.

Item Type: Paper
Uncontrolled Keywords: BIOCHEMICAL-CHARACTERIZATION MAMMALIAN HOMOLOG BETA-COP IN-VITRO PROTEIN KINASE SUBSTRATE BAND-4.1 COMPLEX CELLS
Subjects: bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification
organs, tissues, organelles, cell types and functions > organelles, types and functions > endoplasmic reticulum
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein expression > phosphorylation
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein expression
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > enzymes > protein tyrosine phosphatase
CSHL Authors:
Communities: CSHL labs > Tonks lab
Depositing User: Matt Covey
Date: December 2000
Date Deposited: 18 Dec 2013 19:52
Last Modified: 18 Dec 2013 19:52
PMCID: PMC17628
Related URLs:
URI: http://repository.cshl.edu/id/eprint/29023

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