The complete sequence of a full length cDNA for human liver glyceraldehyde-3-phosphate dehydrogenase: evidence for multiple mRNA species

Arcari, Paolo; Martinelli, Rosanna; Salvatore, Francesco

doi:10.1093/nar/12.23.9179

Cited by 392 publications

(134 citation statements)

References 25 publications

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“…Sequencing analysis (Perkin Elmer Corp., PE Applied Biosystems, Foster City, CA, USA) confirmed the identity of the amplified DNA. A cDNA fragment of human glyceraldehyde-3-phosphate dehydrogenase (G3PDH; residues 71±1053) cDNA was cloned as described for IL-12 [16].…”

Section: Il-12 Assaymentioning

confidence: 99%

Regulation of LPS induced IL-12 production by IFN-γand IL-4 through intracellular glutathione status in human alveolar macrophages

Dobashi

Aihara

Araki

et al. 2001

Clinical and Experimental Immunology

112

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SUMMARYInterleukin-12 (IL-12) is secreted from monocytes and macrophages; it exerts pleiotropic effects on T cells and natural killer (NK) cells, and stimulates interferon-g (IFN-g ) secretion. Glutathione tripeptide regulates the intracellular redox status and other aspects of cell physiology. We examined whether IFNg and IL-4 affect the balance between intracellular reduced glutathione (GSH) and oxidized (GSSG) glutathione, as this may affect IL-12 production in human alveolar macrophages (AM). We used both AM from healthy non-smokers obtained by bronchoalveolar lavage and the monocytic THP-1 cell line in this study. Incubation of AM for 2 h with the GSH precursor N-acetylcysteine (NAC) increased the intracellular GSH/GSSG ratio, and enhanced lipopolysaccharide (LPS)-induced IL-12 secretion by AM. In THP-1 cells, NAC increased the GSH/GSSG ratio and the expression of LPS-induced IL-12 mRNA, whereas l-buthionine-[S,R]-sulphoximine (BSO) decreased these. NAC and BSO offset their own effects on the intracellular GSH/GSSG ratio and the expression of LPS-induced IL-12 mRNA. Furthermore, exposure of AM to the helper T cell type 1 (Th1) cytokine IFN-g or the helper T cell type 2 (Th2) cytokine IL-4 for 72 h increased and decreased the GSH/GSSG ratio, respectively. Lipopolysaccharide (LPS)-induced secretion of IL-12 in AM was enhanced by IFN-g but inhibited by IL-4. These results suggest that IFN-g and IL-4 oppositely affect the GSH/GSSG balance, which may regulate IL-12 secretion from AM in response to LPS.

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Section: Il-12 Assaymentioning

confidence: 99%

Regulation of LPS induced IL-12 production by IFN-γand IL-4 through intracellular glutathione status in human alveolar macrophages

Dobashi

Aihara

Araki

et al. 2001

Clinical and Experimental Immunology

112

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“…PCR was preceded by 2 min at 95°C and followed by 7 min at 72°C. GAPDH primers 26 were used as internal control.…”

Section: Rt-pcr Amplificationmentioning

confidence: 99%

Inhibition of tumor growth and metastatic spreading by overexpression of inter‐alpha‐trypsin inhibitor family chains

Paris

Sesboüé

Delpech³

et al. 2002

Intl Journal of Cancer

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The inter-␣ trypsin inhibitor (ITI) family is a group of proteins built up from different combinations of 1 light chain (ITI-L) and 3 highly homologous heavy chains (ITI-H1, -H2 and -H3). To investigate a potential role of the ITI family chains in cancer and metastasis spreading, we engineered human H460M cell lines expressing both the green fluorescent protein (GFP) and one of these chains. These clones were subcutaneously injected in athymic nude mice, and lung metastasis number and primary tumor weight were determined after 28 days. Expression of the ITI-L chain considerably decreased tumor weight and fluorescent lung metastasis number. ITI-H1 and ITI-H3 chain expression induced a significant decrease of metastasis number, whereas no decrease of tumor weight could be detected. Key words: inter-␣-trypsin inhibitor; green fluorescent protein; spontaneous lung metastases; primary tumorInhibition of the metastatic process represents an important challenge in cancer treatment, and potential therapeutic approaches include the inhibition of matrix metalloproteinases, 1 of the vascular endothelial cell growth factor (VEGF) pathway 2 or the activation of metastasis suppressor genes. 3 Although most of the molecules involved in the metastatic process have not been identified, it is now established that alteration of the extracellular matrix (ECM) plays an important role in cell migration and metastasis. 4 The major ECM component, hyaluronic acid (HA), is organized as a complex network of macromolecules involving hyaluronic acidbinding proteins (HABP) 5 and the ITI family. The most of ITI members are composed of variable associations of light (ITI-L) and heavy chains (ITI-H1, -H2, -H3), encoded by 4 different genes on 3 chromosomes, 6 -9 giving rise to a series of proteins covalently linked by a glycosaminoglycan 10 and differing in their chain composition. 11 Although the binding of ITI to HA is known for a long time, 12 involvement of heavy chains in this linkage was only recently demonstrated 13-15 together with a major role in the ECM stability of the mouse cumulus oocyte complex. 16 The ITI-L chain mRNA encodes 2 tandemly arranged proteins that separate after proteolytic cleavage: the ␣ 1 -microglobulin (␣ 1 m) that belongs to the lipocalin superfamily 17 and the bikunin that contains 2 Kunitz inhibitory domains responsible for the protease inhibitory activity of ITI molecules. 18 Different studies have suggested a potential antimetastatic effect of bikunin: (i) it inhibits in vitro tumor cell receptor-bound plasmin 19 and the invasiveness of malignant cells; 20,21 and (ii) injections of human purified bikunin efficiently reduced the number of metastases produced by the murine 3LL cell line. 22 We have therefore analyzed in the present study the antimetastatic effect of the different ITI chains using a model of fluorescent spontaneous lung metastases. 23 MATERIAL AND METHODS Cells lines and cultureThe H460M cell line, initially derived from the parental NCI-H460 cell line established from a large-cell carcinoma o...

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“…Cytoplasmic RNA was extracted from cells using Trizol reagent (Gibco BRL). RNA (2 mg) was resolved on formaldehyde agarose gel, transferred to nylon membrane and hybridized to 32 P-labeled EF-1A cDNA (clone pAN7) or 32 P-labeled GAPDH cDNA 30 probes as described by Maniatis et al 31 Cytosolic extracts and Western blot analysis. H1355 cells were scraped, washed twice in cold phosphate-buffered saline (PBS) and resuspended in 20-40 ml of lysis buffer (50 mM Tris-HCl pH 7.4, 1% NP40, 0.25% sodium deoxycholate, 150 mM NaCl, 1 mg/ml aprotinin, leupeptin, pepstatin, 1 mM Na 3 VO 4 , 1 mM NaF) for 30 min on ice and centrifuged at 14 000 Â g for 20 min at 41C.…”

Section: Methodsmentioning

confidence: 99%

C-Raf antagonizes apoptosis induced by IFN-α in human lung cancer cells by phosphorylation and increase of the intracellular content of elongation factor 1A

Lamberti

Longo

Marra³

et al. 2007

Cell Death Differ

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Interferon a (IFNa) induces both apoptosis and a counteracting epidermal growth factor Erk-dependent survival response in cancer cells. In this report, IFNa increased eukaryotic elongation factor 1A (eEF-1A) protein expression by inhibition of eEF-1A degradation via a proteasome-dependent pathway. The reduction of the expression level of eEF-1A by RNA interference enhanced the apoptosis induced by IFNa on the same cells. Moreover, IFNa induced the phosphorylation of both serine and threonine in eEF-1A. These effects were paralleled by an increased co-immunoprecipitation and colocalization of eEF-1A with C-Raf. The suppression of C-Raf kinase activity with the inhibitor BAY 43-9006 completely antagonized the increase of both eEF-1A phosphorylation and expression and of C-Raf/eEF-1A colocalization induced by IFNa and enhanced apoptosis and eEF-1A ubiquitination. Cell transfection with the mutated K48R ubiquitin increased EF-1A expression and desensitized tumor cells to the modulating effects of IFNa. The dynamic simulation of 3Dstructure of eEF-1A identified putative serine and threonine phosphorylation sites. In conclusion, the interaction between eEF-1A and C-Raf increases eEF-1A stability and induces a survival activity.

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The complete sequence of a full length cDNA for human liver glyceraldehyde-3-phosphate dehydrogenase: evidence for multiple mRNA species

Cited by 392 publications

References 25 publications

Regulation of LPS induced IL-12 production by IFN-γand IL-4 through intracellular glutathione status in human alveolar macrophages

Regulation of LPS induced IL-12 production by IFN-γand IL-4 through intracellular glutathione status in human alveolar macrophages

Inhibition of tumor growth and metastatic spreading by overexpression of inter‐alpha‐trypsin inhibitor family chains

C-Raf antagonizes apoptosis induced by IFN-α in human lung cancer cells by phosphorylation and increase of the intracellular content of elongation factor 1A

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