GD3 Synthase Gene Expression in PC12 Cells Results in the Continuous Activation of TrkA and ERK1/2 and Enhanced Proliferation

Fukumoto, Satoshi; Mutoh, Tatsuro; Hasegawa, Tomokazu; Miyazaki, Hiroshi; Okada, Masahiko; Goto, G; Furukawa, Koichi; Urano, Takeshi

doi:10.1074/jbc.275.8.5832

Cited by 107 publications

(68 citation statements)

References 41 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Whereas CHO-GD3 cells express mainly GM3, GD3, and 9-O-acetyl-GD3, transfection of Neuro2a cells with the GD3 synthase gene induces not only GD3 but also other larger b-series gangliosides (74). Similar findings were observed in rat pheochromocytoma PC12 cells, in which transfection of the GD3 synthase gene resulted in drastic increase of GD1b and GT1b (75). Thus, the outcome of GD3 synthase expression could depend not only upon the cell type but also upon the preexisting ganglioside profile in the cells.…”

Section: Tis21 and 9-o-acetylation Of Gd3supporting

confidence: 67%

Genes Modulated by Expression of GD3 Synthase in Chinese Hamster Ovary Cells

Satake¹,

Chen²,

Varki³

2003

Journal of Biological Chemistry

View full text Add to dashboard Cite

9-O-Acetylation is a common sialic acid modification, expressed in a developmentally regulated and tissue/ cell type-specific manner. The relevant 9-O-acetyltransferase(s) have not been isolated or cloned; nor have mechanisms for their regulation been elucidated. We previously showed that transfection of the GD3 synthase (ST8Sia-I) gene into Chinese hamster ovary (CHO)-K1 cells gave expression of not only the disialoganglioside GD3 but also 9-O-acetyl-GD3. We now use differential display PCR between wild type CHO-K1 cells and clones stably expressing GD3 synthase (CHO-GD3 cells) to detect any increased expression of other genes and explore the possible induction of a 9-O-acetyltransferase. The four CHO mRNAs showing major upregulation were homologous to VCAM-1, Tis21, the KCprotein-like protein, and a functionally unknown type II transmembrane protein. A moderate increase in expression of the FxC1 and SPR-1 genes was also seen. Interestingly, these are different from genes observed by others to be up-regulated after transfection of GD3 synthase into a neuroblastoma cell line. We also isolated a CHO-GD3 mutant lacking 9-O-acetyl-GD3 following chemical mutagenesis (CHO-GD3-OAc ؊ ). Analysis of the above differential display PCR-derived genes in these cells showed that expression of Tis21 was selectively reduced. Transfection of a mouse Tis21 cDNA into the CHO-GD3-OAc ؊ mutant cells restored 9-O-acetyl-GD3 expression. Since the only major gangliosides expressed by CHO-GD3 cells are GD3 and 9-O-acetyl-GD3 (in addition to GM3, the predominant ganglioside type in wildtype CHO-K1 cells), we conclude that GD3 enhances its own 9-O-acetylation via induction of Tis21. This is the first known nuclear inducible factor for 9-O-acetylation and also the first proof that 9-O-acetylation can be directly regulated by GD3 synthase. Finally, transfection of CHO-GD3-OAc ؊ mutant cells with ST6Gal-I induced 9-O-acetylation specifically on sialylated N-glycans, in a manner similar to wild-type cells. This indicates separate machineries for 9-O-acetylation on ␣2-8-linked sialic acids of gangliosides and on ␣2-6-linked sialic acids on N-glycans.Sialic acids are a family of 9-carbon carboxylated monosaccharides typically located at the termini of mammalian cell surface sugar chains on both glycoproteins and glycolipids. N-acetylneuraminic acid, the most common sialic acid, is subjected to various modifications in vivo (1-5). One of the most prevalent modifications is O-acetylation of the hydroxyl group at the 9-carbon position. This modification is known to reduce or abolish the recognition of sialic acid residues by sialidases (2, 4, 6, 7), by certain sialic acid-binding lectins like Siglecs (4, 5, 8 -10), and by several viral recognition proteins (4,5,11,12). Conversely, other viruses require 9-O-acetylation for recognition of their target cells (4,5,(13)(14)(15)(16). It is also known that 9-O-acetylation is regulated during development and aberrantly expressed in melanomas and basal cell carcinomas (17)(18)(19)(20). These findings ind...

show abstract

Section: Tis21 and 9-o-acetylation Of Gd3supporting

confidence: 67%

Genes Modulated by Expression of GD3 Synthase in Chinese Hamster Ovary Cells

Satake¹,

Chen²,

Varki³

2003

Journal of Biological Chemistry

View full text Add to dashboard Cite

show abstract

“…These gangliosides also modulate immunoglobulin synthesis in peripheral blood mononuclear cells Tamaki, 1998, 1999a, b) and cause a shift from Th-2 to Th-1 cytokine production in PHA-stimulated T cells . Finally, increased expression of GD1b and GT1b in rat pheochromocytoma cells transfected with GD3 synthase occurs in parallel with trk-A dimerisation (Fukumoto et al, 2000), which in turn is associated with improved prognosis in NB (Nakagawara et al, 1993). Taking these findings together, we hypothesise that CbG may impede tumour progression, either by acting directly on the tumour cell or by increasing the host resistance.…”

Section: Discussionmentioning

confidence: 85%

“…We have linked alterations in the expression of complex 'b' pathway gangliosides (CbG) downstream of GD1b/GM1a synthase (GD1b, GT1b and GQ1b) to differences in the biological phenotype and clinical behaviour that can be used to predict patient outcome in NB (Hettmer et al, 2003). Further, recent experimental evidence suggests that GD1b, GT1b and GQ1b can modulate a number of biological processes that are believed to counteract malignant transformation and progression, including tumour cell proliferation, host immune function and signal transduction mechanisms (Hynds et al, 1995;Fukumoto et al, 2000;. We hypothesise that high CbG content may contribute to reduced tumour aggressiveness, and that alteration of CbG expression might be a potential therapeutic target in NB.…”

mentioning

confidence: 92%

Alterations in neuroblastoma ganglioside synthesis by induction of GD1b synthase by retinoic acid

et al. 2004

View full text Add to dashboard Cite

Recent findings link increased expression of the structurally complex 'b' pathway gangliosides GD1b, GT1b, GQ1b (CbG) to a favourable clinical and biological behaviour in human neuroblastoma (NB). Seeking a model to probe these observations, we evaluated four human NB cell lines. Very low CbG content (4 -10%) in three of the four cell lines (LAN-5, LAN-1, SMS-KCNR) reflected the ganglioside pattern observed in the most aggressive NB tumours. Pharmacological alterations of complex ganglioside synthesis in vitro by a 5 -7 day exposure to 5 -10 mM retinoic acid, which is employed in maintenance therapy of disseminated NB, included markedly increased (i) relative expression of CbG (6.672.0-fold increase, P ¼ 0.037), (ii) relative expression of the analogous 'a' pathway gangliosides, termed CaG (6.471.4-fold increase in GM1a and GD1a; P ¼ 0.010), and (iii) total cellular ganglioside content (2.0 -6.3-fold), which in turn amplified the accumulation of structurally complex gangliosides. Substantial increases (2.7 -2.9-fold) in the activity of GD1b/GM1a synthase (b-1,3-galactosyltransferase), which initiates the synthesis of CbG and CaG, accompanied the all-trans retinoic acid (ATRA)-induced ganglioside changes. Thus, increased CbG synthesis in NB cell lines is attributable to a specific effect of ATRA, namely induction of GD1b/GM1a synthase activity. Since the shift towards higher expression of CbG and CaG during retinoic acid-induced cellular differentiation reflects a ganglioside pattern found in clinically less-aggressive tumours, our studies suggest that complex gangliosides may play a role in the biological and clinical behaviour of NB.

show abstract

“…We have also suggested the possibility that ganglioside GM2 modulates integrin function (47). Simultaneously, GD3 might also interact with unidentified growth factor receptor(s) and enhance the receptor function, as we demonstrated in the study of a rat pheochromocytoma cell line, PC12 (48). The elucidation of molecules associating with GD3 in GD3 ϩ cells is critical.…”

Section: Discussionmentioning

confidence: 95%

Ganglioside GD3 promotes cell growth and invasion through p130Cas and paxillin in malignant melanoma cells

Hamamura

Hayashi²,

Hattori³

et al. 2005

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

146

163

View full text Add to dashboard Cite

Although ganglioside GD3 levels are highly elevated in malignant melanomas, the role of GD3 in melanomas' malignant properties has not been clearly shown. To investigate this problem, we genetically generated GD3-positive (GD3 ؉ ) transfectant cells from a GD3-negative (GD3 ؊ ) mutant line SK-MEL-28-N1 and analyzed the phenotypic changes in the transfected cells. GD3 ؉ cells showed markedly increased cell growth and invasive characteristics. Two bands that underwent stronger tyrosine phosphorylation in GD3 ؉ cell lines than in controls after treatment with FCS were found with molecular masses of 130 and 68 kDa. They were identified as p130Cas and paxillin by sequential immunoprecipitation. Their roles in cell growth and invasion were analyzed with a small interfering RNA (siRNA) approach. Cell growth, as analyzed by BrdUrd uptake, was strongly suppressed in GD3 ؉ cells to near the levels of GD3 ؊ cells when treated with siRNA for p130Cas but not when treated with siRNA for paxillin. However, treatment with siRNAs of either p130Cas or paxillin resulted in the marked suppression of the invasive activity of GD3 ؉ cells almost to the levels of control cells. These results suggested that these two molecules function as effectors of GD3-mediated signaling, leading to such malignant properties as rapid cell growth and invasion.small interfering RNA ͉ sialyltransferase ͉ phosphorylation

show abstract

GD3 Synthase Gene Expression in PC12 Cells Results in the Continuous Activation of TrkA and ERK1/2 and Enhanced Proliferation

Cited by 107 publications

References 41 publications

Genes Modulated by Expression of GD3 Synthase in Chinese Hamster Ovary Cells

Genes Modulated by Expression of GD3 Synthase in Chinese Hamster Ovary Cells

Alterations in neuroblastoma ganglioside synthesis by induction of GD1b synthase by retinoic acid

Ganglioside GD3 promotes cell growth and invasion through p130Cas and paxillin in malignant melanoma cells

Contact Info

Product

Resources

About