Document Detail


Atypical protein kinase C plays a critical role in protein transport from pre-Golgi intermediates.
MedLine Citation:
PMID:  12871960     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The small GTPase Rab2 requires atypical protein kinase C iota/lambda (PKCiota/lambda) kinase activity to promote vesicle budding from normal rat kidney cell microsomes (Tisdale, E. J. (2000) Traffic 1, 702-712). The released vesicles lack anterograde-directed cargo but contain coat protein I (COPI) and the recycling protein p53/p58, suggesting that the vesicles traffic in the retrograde pathway. In this study, we have directly characterized the role of PKCiota/lambda in the early secretory pathway. A peptide corresponding to the unique PKCiota/lambda pseudosubstrate domain was introduced into an in vitro assay that efficiently reconstitutes transport of vesicular stomatitis virus glycoprotein from the endoplasmic reticulum to the cis-medial Golgi compartments. This peptide blocked transport in a dose-dependent manner. Moreover, normal rat kidney cells incubated with Rab2 and the pseudosubstrate peptide displayed abundant swollen or dilated vesicles that contained Rab2, PKCiota/lambda, beta-COP, and p53/p58. Because Rab2, beta-COP, and p53/p58 are marker proteins for pre-Golgi intermediates (vesicular tubular clusters,VTCs), most probably the swollen vesicles are derived from VTCs. Similar results were obtained when the assays were supplemented with kinase-dead PKCiota/lambda (W274K). Both the pseudosubstrate peptide and kinase-dead PKCiota/lambda in tandem with Rab2 caused sustained membrane association of PKCiota/lambda, suggesting that reverse translocation was inhibited. Importantly, the inhibitory phenotype of kinase-dead PKCiota/lambda was reversed by PKCiota/lambda wild type. These combined results indicate that PKCiota/lambda is essential for protein transport in the early secretory pathway and suggest that PKCiota/lambda kinase activity is required to promote Rab2-mediated vesicle budding at a VTC subcompartment enriched in recycling cargo.
Authors:
Ellen J Tisdale; Jing Wang; Robert B Silver; Cristina R Artalejo
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S.; Research Support, U.S. Gov't, P.H.S.     Date:  2003-07-17
Journal Detail:
Title:  The Journal of biological chemistry     Volume:  278     ISSN:  0021-9258     ISO Abbreviation:  J. Biol. Chem.     Publication Date:  2003 Sep 
Date Detail:
Created Date:  2003-09-22     Completed Date:  2003-11-17     Revised Date:  2012-06-13    
Medline Journal Info:
Nlm Unique ID:  2985121R     Medline TA:  J Biol Chem     Country:  United States    
Other Details:
Languages:  eng     Pagination:  38015-21     Citation Subset:  IM    
Affiliation:
Department of Pharmacology, Wayne State University School of Medicine, Detroit, Michigan 48201, USA. etisdale@med.wayne.edu
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MeSH Terms
Descriptor/Qualifier:
Amino Acid Sequence
Animals
Endoplasmic Reticulum / metabolism
Golgi Apparatus / metabolism*
Isoenzymes / physiology*
Kidney / metabolism*,  ultrastructure*
Membrane Glycoproteins / metabolism
Microsomes / metabolism
Molecular Sequence Data
Protein Kinase C / physiology*
Protein Transport
Rats
Viral Envelope Proteins / metabolism
rab2 GTP-Binding Protein / physiology
Grant Support
ID/Acronym/Agency:
DK58921/DK/NIDDK NIH HHS
Chemical
Reg. No./Substance:
0/G protein, vesicular stomatitis virus; 0/Isoenzymes; 0/Membrane Glycoproteins; 0/Viral Envelope Proteins; EC 2.7.11.13/Protein Kinase C; EC 2.7.11.13/protein kinase C lambda; EC 3.6.5.2/rab2 GTP-Binding Protein

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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