Abstract
Alcohol dehydrogenase-I (ADH-I) derived from horse liver stimulated IgM production by human-human hybridoma, HB4C5 cells and lymphocytes. The IPSF activity of ADH-I was suppressed by coexistence of short DNA whose chain length is less than 200 base pairs (bp) and fibrous DNA in a dose-dependent manner. These DNA preparations completely inhibited the IPSF activity at the concentration of 250 μg/ml and 1.0 mg/ml, respectively. DNA sample termed long DNA whose average chain length is 400–7000 bp slightly stimulated IPSF activity at 0.06 μg/ml. However, long DNA suppressed IPSF activity by half at 1.0 mg/ml. The laser confocal microscopic analysis had revealed that ADH-I was incorporated by HB4C5 cells. The uptake of ADH-I was strongly inhibited by short DNA and fibrous DNA. However, long DNA did not suppress the internalization of ADH-I into HB4C5 cells. These findings indicate that short DNA and fibrous DNA depress IPSF activity of ADH-I by inhibiting the internalization of this enzyme. According to the gel-filtration analysis using HPLC, ADH-I did not directly interact with short DNA. It is expected from these findings that short DNA influences HB4C5 cells to suppress the internalization of ADH-I. Moreover, these facts also strongly suggest that ADH-I acts as IPSF after internalization into the cell.
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Miyazaki Y, Nishimoto S, Sasaki T and Sugahara T (1998) Spermine enhances IgM productivity of human-human hybridoma HB4C5 cells and human peripheral blood lymphocytes. Cytotechnology 26: 111–118.
Murakami H, Masui H, Sato GH, Sueoka N, Chow TP and Kono T (1982) Growth of hybridoma cells in serum-free medium. Proc Natl Acad Sci USA 79: 1158–1162.
Murakami H, Hashizume S, Ohashi H, Shinohara K, Yasumoto K, Nomoto K and Omura H (1985) Human-human hybridomas secreting antibodies specific to human lung carcinoma. In Vitro Cell Develop Biol 21: 593–596.
Nagy E and Rigby WFC (1995) Glyceraldehyde-3-phosphate dehydrogenase selectively binds AU-rich RNA in the NADC+-binding region (Rossmann fold). J Biol Chem 270: 2755–2763
Perucho M, Salas J and Salas ML (1977) Identification of the mammalian DNA-binding protein P8 as glyceraldehyde-3-phosphate dehydrogenase. Eur J Biochem 81: 557–562.
Perucho M, Salas J and Salas ML (1980) Study of the interaction of glyceraldehyde-3-phosphate dehydrogenase with DNA. Biochim Biophys Acta 606: 181–195.
Ryazanov AG (1985) Glyceraldehyde-3-phosphate dehydrogenase is one of the three major RNA-binding proteins of rabbit reticulocytes. FEBS Lett 192: 131–134.
Ryazanov AG, Ashmarina LI and Muronetz VI (1988) Association of glyceraldehyde-3-phosphate dehydrogenase with mono-and polyribosomes of rabbit reticulocytes. Eur J Biochem 171: 301–305.
Sabath DE, Broome HE and Prystowsky MB (1990) Glyceraldehyde-3-phosphate dehydrogenase mRNA is a major interleukin 2-induced transcript in a cloned T-helper lymphocyte. Gene 91: 185–191.
Singh R and Green MR (1995) Sequence-specific binding of transfer RNA by glyceraldehyde-3-phosphate dehydrogenase. Science 259: 365–368.
Sugahara T, Shirahata S, Yamada K and Murakami H (1991) Purification of immunoglobulin production stimulating factor-IIα derived from Namalwa cells. Cytotechnology 5: 255–263.
Sugahara T, Sasaki T and Murakami H (1994) Enhancement of immunoglobulin productivity of human-human hybridoma HB4C5 cells by basic proteins and poly-basic amino acids. Biosci Biotech Biochem 58: 2212–2214.
Sugahara T, Shirahata S, Sasaki T and Murakami H (1995) The mode of actions of glyceraldehyde-3-phosphate dehydrogenase identified as an immunoglobulin production stimulating factor. FEBS Lett 368: 92–96.
Sugahara T, Furutani H and Sasaki T (1997) Alcohol dehydrogenase-I from horse liver stimulates immunoglobulin production by human hybridoma and human peripheral blood lymphocytes. Mol Cell Biochem 173: 113–119.
Sugahara T and Sasaki T (1998) Inhibition of immunoglobulin production stimulating activity of glyceraldehyde-3-phosphate dehydrogenase by nucleotides. Biosci Biotech Biochem 62: 1237–1239.
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Okamoto, T., Furutani, H., Sasaki, T. et al. Effects of DNA on immunoglobulin production stimulating activity of alcohol dehydrogenase. Cytotechnology 31, 95–102 (1999). https://doi.org/10.1023/A:1008024322602
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DOI: https://doi.org/10.1023/A:1008024322602