Document Detail


Characterization of the functional domains of galactosylceramide expression factor 1 in MDCK cells.
MedLine Citation:
PMID:  11555619     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
We previously reported that GalCer expression factor 1 (GEF-1), a rat homologue of hepatocyte growth factor-regulated tyrosine kinase substrate (Hrs), induced GalCer expression, morphological changes, and cell growth inhibition in COS-7 cells. In this study, we describe the characterization of GEF-1 in MDCK cells. Overexpression of GEF-1 in MDCK (MDCK/GEF-1) cells showed GalCer-derived sulfatide expression as well as dramatic morphological changes, but not cell growth suppression. The enzyme activity and the mRNA level of UDP-galactose:ceramide galactosyltransferase (CGT) increased significantly in MDCK/GEF-1 cells compared with control cells. GEF-1 molecule is composed of four domains; a zinc-finger (Z), a proline-rich (P), a coiled-coil (C), and a proline/glutamine-rich (Q) domain. MDCK cells transfected with various GEF-1 deletion mutants were examined for morphology and for glycolipid expression. MDCK cells transfected with Z-domain deletion mutant (MDCK/PCQ) and those with both Z- and P-domains deletion mutant (MDCK/CQ) were similar to those with a wild-type GEF-1 (MDCK/ZPCQ) in shape, exhibiting fibroblast-like cells, whereas those with the other deletion mutants showed no morphological changes, exhibiting typical epithelial-like cells. On the other hand, MDCK/ZPCQ, MDCK/PCQ, MDCK/CQ, and MDCK/Q cells expressed sulfatide, whereas those with the other deletion mutants that did not include the Q-domain showed neither GalCer nor sulfatide expression. Thus, the correlation between fibroblast-like cells in shape and the glycolipid expression was good in these deletion mutants except MDCK/Q cells, which showed epithelial-like cells, but expressed sulfatide. The glycolipid expression paralleled CGT mRNA levels. Taking these results together, it is suggested that only the Q-domain may be essential for the role of GEF-1 in inducing CGT mRNA, whereas the Q-domain together with the C-domain may be required for the induction of morphological changes in MDCK cells.
Authors:
K Ogura; T Tai
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Glycobiology     Volume:  11     ISSN:  0959-6658     ISO Abbreviation:  Glycobiology     Publication Date:  2001 Sep 
Date Detail:
Created Date:  2001-09-13     Completed Date:  2001-12-07     Revised Date:  2009-11-19    
Medline Journal Info:
Nlm Unique ID:  9104124     Medline TA:  Glycobiology     Country:  England    
Other Details:
Languages:  eng     Pagination:  751-8     Citation Subset:  IM    
Affiliation:
Department of Tumor Immunology, The Tokyo Metropolitan Institute of Medical Science, Honkomagome, Bunkyo-ku, Tokyo 113-8613, Japan.
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MeSH Terms
Descriptor/Qualifier:
Adenosine Triphosphatases*
Animals
Base Sequence
Cell Line
Cell Movement / physiology
DNA Primers
DNA, Complementary
Dogs
Electrophoresis, Polyacrylamide Gel
Endosomal Sorting Complexes Required for Transport
Enzyme Inhibitors / pharmacology
Galactosylceramidase / genetics,  metabolism*
Hepatocyte Growth Factor / physiology
Immunohistochemistry
Morphogenesis / physiology
Phosphoproteins / chemistry,  genetics,  metabolism*
Protein-Tyrosine Kinases / antagonists & inhibitors
Sequence Deletion
Transfection
Chemical
Reg. No./Substance:
0/DNA Primers; 0/DNA, Complementary; 0/Endosomal Sorting Complexes Required for Transport; 0/Enzyme Inhibitors; 0/Hgs protein, rat; 0/Phosphoproteins; 67256-21-7/Hepatocyte Growth Factor; EC 2.7.10.1/Protein-Tyrosine Kinases; EC 3.2.1.46/Galactosylceramidase; EC 3.6.1.-/Adenosine Triphosphatases

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


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