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A mutation in glyceraldehyde 3-phosphate dehydrogenase alters endocytosis in CHO cells.
- Source :
-
The Journal of cell biology [J Cell Biol] 1995 Sep; Vol. 130 (5), pp. 1093-104. - Publication Year :
- 1995
-
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.
- Subjects :
- Animals
Base Sequence
CHO Cells metabolism
CHO Cells ultrastructure
Cell Compartmentation genetics
Cloning, Molecular
Cricetinae
DNA, Complementary genetics
Fluorescent Dyes
Gene Expression physiology
Glyceraldehyde-3-Phosphate Dehydrogenases genetics
Microscopy, Electron
Microtubule-Associated Proteins genetics
Molecular Sequence Data
Paclitaxel metabolism
Peptide Fragments genetics
Phenotype
Protein Binding physiology
Endocytosis genetics
Glyceraldehyde-3-Phosphate Dehydrogenases metabolism
Peptide Fragments metabolism
Point Mutation physiology
Subjects
Details
- Language :
- English
- ISSN :
- 0021-9525
- Volume :
- 130
- Issue :
- 5
- Database :
- MEDLINE
- Journal :
- The Journal of cell biology
- Publication Type :
- Academic Journal
- Accession number :
- 7657694
- Full Text :
- https://doi.org/10.1083/jcb.130.5.1093