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PMID: 14617820 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Distinct roles for the AAA ATPases NSF and p97 in the secretory pathway.

Molecular biology of the cell ·Vol. 15 ·No. 2 ·2004-02-00 ·Pages 637-48

Dalal S, Rosser MF, Cyr DM, Hanson PI

Abstract

NSF and p97 are related AAA proteins implicated in membrane trafficking and organelle biogenesis. p97 is also involved in pathways that lead to ubiquitin-dependent proteolysis, including ER-associated degradation (ERAD). In this study, we have used dominant interfering ATP-hydrolysis deficient mutants (NSF(E329Q) and p97(E578Q)) to compare the function of these AAA proteins in the secretory pathway of mammalian cells. Expressing NSF(E329Q) promotes disassembly of Golgi stacks into dispersed vesicular structures. It also rapidly inhibits glycosaminoglycan sulfation, reflecting disruption of intra-Golgi transport. In contrast, expressing p97(E578Q) does not affect Golgi structure or function; glycosaminoglycans are normally sulfated and secreted, as is the VSV-G ts045 protein. Instead, expression of p97(E578Q) causes ubiquitinated proteins to accumulate on ER membranes and slows degradation of the ERAD substrate cystic-fibrosis transmembrane-conductance regulator. In addition, expression of p97(E578Q) eventually causes the ER to swell. More specific assessment of effects of p97(E578Q) on organelle assembly shows that the Golgi apparatus disperses and reassembles normally after treatment with brefeldin A and during mitosis. These findings demonstrate that ATP-hydrolysis-dependent activities of NSF and p97 in the cell are not equivalent and suggest that only NSF is directly involved in regulating membrane fusion.

MeSH Terms
Adenosine Triphosphatases/metabolism Animals Brefeldin A/pharmacology Cells, Cultured Cloning, Molecular Endoplasmic Reticulum/metabolism Glycosaminoglycans/metabolism Golgi Apparatus/metabolism Green Fluorescent Proteins Humans Intracellular Membranes/metabolism Luminescent Proteins/metabolism Membrane Fusion/drug effects,physiology Microscopy, Electron Mutation/genetics Nuclear Proteins/metabolism Protein Structure, Tertiary/physiology
Chemicals
Glycosaminoglycans Luminescent Proteins Nuclear Proteins Green Fluorescent Proteins Brefeldin A Adenosine Triphosphatases p97 ATPase
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Dalal Seema
Washington University School of Medicine, Department of Cell Biology and Physiology, St. Louis, Missouri 63110, USA.
Rosser Meredith F N
Cyr Douglas M
Hanson Phyllis I
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Article Info
Journal
Molecular biology of the cell
Abbr.
Mol Biol Cell
ISSN
1059-1524
DOI
10.1091/mbc.e03-02-0097
Published
2004-02-00
Epub
2003-00-14
Pages
637-48
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC329284
Subset
IM
Grants
NINDS NIH HHS · F32 NS041709 · United States
NINDS NIH HHS · NS38058 · United States
NIGMS NIH HHS · R01 GM056981 · United States
NINDS NIH HHS · NS41709 · United States
NINDS NIH HHS · R01 NS038058 · United States
External Links
PubMed source DOI: 10.1091/mbc.e03-02-0097 PMC Full Text View journal details
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