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

Abstract

Abstract. The CHO cell mutant FD1.3.25 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 ~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 ~15% of the total enzyme exhibited the various changes in endocytosis observed in FD1.3.25.

Flocculation of Saccharomyces cerevisiae is induced by transformation with the GAP1 gene from Kluyveromyces marxianus

“…The ubiquity of GAPDH has been the subject of several reports in the literature (Huitorel and Pantolini, 1985;Kawamoto and Caswell, 1986;Allen et al, 1987;Meyer-Siegler et al, 1991;Pancholi and Fischetti, 1992;Singh and Green, 1993;Robbins et al, 1995;GilNavarro et al, 1997) and the results presented in this paper clearly show that this GAPDH protein (p37) has other functions than its glycolytic enzymatic activity, as its oversynthesis in¯uences growth, biomass yield, cell wall stability, cell morphology and aggregation. There is now evidence that p37 can induce cell aggregation in Saccharomyces, although it is a Kluyveromyces protein.…”

Identification of glyceraldehyde-3-phosphate dehydrogenase as a Ca2+-dependent fusogen in human neutrophil cytosol

“…We also detected low levels of Ca 2ϩ -independent aggregation and fusion activities, but this accounted for only a small fraction of that achievable with Ca 2ϩ (not shown). We have also shown that GAPDH in our fusion system apparently has a potential for Ca 2ϩ -mediated fusion that is at least equivalent to annexin I. Robbins et al [28] recently discovered in late endosomes that a single amino acid substitution in GAPDH caused a marked difference in endocytosis of liquid phase markers. Thus, it can be seen in an unrelated system that GAPDH appears to serve in a membrane-membrane fusion capacity.…”

Glyceraldehyde 3-phosphate dehydrogenase is a SET-binding protein and regulates cyclin B-cdk1 activity

“…Interestingly, it is clear now that GAPDH displays a distinct membrane, cytosolic and nuclear localization. In those subcellular sites, it is involved in a number of cellular functions, including endocytosis and membrane fusion (Glaser and Gross, 1995;Robbins et al, 1995), vesicular secretory transport (Tisdale, 2001), translational control (Yi et al, 2000), nuclear tRNA transport (Singh and Green, 1993), DNA replication (Zheng et al, 2003) and DNA repair (Krynetski et al, 2001).…”