Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
4
pubmed:dateCreated
2002-1-21
pubmed:abstractText
Recent work from our laboratory demonstrated that phosphatidic acid (PA) and phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P(2)), are required to maintain the structural integrity of the Golgi apparatus. To investigate the role of these lipids in regulating Golgi structure and function, we developed a novel assay to follow the release of post-Golgi vesicles. Isolated rat liver Golgi membranes were incubated with [(3)H]CMP sialic acid to radiolabel endogenous soluble and membrane glycoproteins present in the late Golgi and trans-Golgi network. The release of post-Golgi secretory vesicles was determined by measuring incorporation of (3)H-labeled proteins into a medium speed supernatant. Vesicle budding was dependent on temperature, cytosol, energy and time. Electron microscopy of Golgi fractions prior to and after incubation demonstrated that the stacked Golgi cisternae generated a heterogeneous population of vesicles (50- to 350-nm diameter). Inhibition of phospholipase D-mediated PA synthesis, by incubation with 1-butanol, resulted in the complete fragmentation of the Golgi membranes in vitro into 50- to 100-nm vesicles; this correlated with diminished PtdIns(4,5)P(2) synthesis. Following alcohol washout, PA synthesis resumed and in the presence of cytosol PtdIns(4,5)P(2) synthesis was restored. Most significantly, under these conditions the fragmented Golgi elements reformed into flattened cisternae and the re-assembled Golgi supported vesicle release. These data demonstrate that inositol phospholipid synthesis is essential for the structure and function of the Golgi apparatus.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Jan
pubmed:issn
0021-9258
pubmed:author
pubmed:issnType
Print
pubmed:day
25
pubmed:volume
277
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
3030-9
pubmed:dateRevised
2007-11-14
pubmed:meshHeading
pubmed-meshheading:11704660-Alcohols, pubmed-meshheading:11704660-Animals, pubmed-meshheading:11704660-Blotting, Western, pubmed-meshheading:11704660-Brain, pubmed-meshheading:11704660-Cytosol, pubmed-meshheading:11704660-DNA Fragmentation, pubmed-meshheading:11704660-Electrophoresis, Polyacrylamide Gel, pubmed-meshheading:11704660-Glycoproteins, pubmed-meshheading:11704660-Golgi Apparatus, pubmed-meshheading:11704660-Liver, pubmed-meshheading:11704660-Microscopy, Electron, pubmed-meshheading:11704660-N-Acetylneuraminic Acid, pubmed-meshheading:11704660-Phosphatidylinositol 4,5-Diphosphate, pubmed-meshheading:11704660-Phospholipase D, pubmed-meshheading:11704660-Precipitin Tests, pubmed-meshheading:11704660-Rats, pubmed-meshheading:11704660-Structure-Activity Relationship, pubmed-meshheading:11704660-Temperature, pubmed-meshheading:11704660-Time Factors
pubmed:year
2002
pubmed:articleTitle
Fragmentation and re-assembly of the Golgi apparatus in vitro. A requirement for phosphatidic acid and phosphatidylinositol 4,5-bisphosphate synthesis.
pubmed:affiliation
Department of Developmental and Molecular Biology, Albert Einstein College of Medicine, Bronx, New York 10461, USA.
pubmed:publicationType
Journal Article, In Vitro, Research Support, U.S. Gov't, P.H.S.