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PMID: 7657694 Published · ppublish English Journal Article

A mutation in glyceraldehyde 3-phosphate dehydrogenase alters endocytosis in CHO cells.

The Journal of cell biology ·Vol. 130 ·No. 5 ·1995-09-00 ·Pages 1093-104

Robbins AR, Ward RD, Oliver C

Abstract

The CHO cell mutant FD 1.3.25 exhibits both increased accumulation and altered distribution of endocytosed fluid phase tracers. Neither the rate of tracer internalization nor the kinetics of recycling from early endosomes was affected, but exocytosis from late endocytic compartments appeared to be decreased in the mutant. Endocytosed tracer moved more rapidly to the cell poles in FD1.3.25 than in wild type cells. An abundant 36-kD polypeptide was found associated with taxol-polymerized microtubules in preparations from wild type and mutant; in the former but not the latter this polypeptide could be dissociated by incubation of the microtubules in ATP or high salt. The 36-kD polypeptide co-electrophoresed in two dimensions with the monomer of the glycolytic enzyme glyceraldehyde 3-phosphate dehydrogenase (GAPDH). Analysis of cDNA clones showed that the mutant is heterozygous for this enzyme, with approximately 25% of the GAPDH RNA containing a single nucleotide change resulting in substitution of Ser for Pro234, a residue that is conserved throughout evolution. Stable transfectants of wild type cells expressing the mutant monomer at approximately 15% of the total enzyme exhibited the various changes in endocytosis observed in FD1.3.25.

MeSH Terms
Animals Base Sequence CHO Cells/metabolism,ultrastructure Cell Compartmentation/genetics Cloning, Molecular Cricetinae DNA, Complementary/genetics Endocytosis/genetics Fluorescent Dyes Gene Expression/physiology Glyceraldehyde-3-Phosphate Dehydrogenases/genetics,metabolism Microscopy, Electron Microtubule-Associated Proteins/genetics Molecular Sequence Data Paclitaxel/metabolism Peptide Fragments/genetics,metabolism Phenotype Point Mutation/physiology Protein Binding/physiology
Chemicals
DNA, Complementary Fluorescent Dyes Microtubule-Associated Proteins Peptide Fragments glyceraldehyde 3-phosphate dehydrogenase (304-313) Glyceraldehyde-3-Phosphate Dehydrogenases Paclitaxel
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Robbins A R
Laboratory of Biochemistry and Metabolism, National Institute of Diabetes, Digestive and Kidney Diseases, National Institute of Dental Research, Bethesda, Maryland 20892, USA.
Ward R D
Oliver C
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.130.5.1093
Published
1995-09-00
Pages
1093-104
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2120546
Subset
IM
External Links
PubMed source DOI: 10.1083/jcb.130.5.1093 PMC Full Text View journal details
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