Abstract
One of the Ser-tRNAs, Ser-tRNASec, is converted to Sec-tRNASec by Sec synthase. This Ser-tRNASec is also converted to phosphoser-tRNASec by tRNA kinase. In this study, we analyzed of the products of phosphorylation with tRNA kinase. [3H]Ser-tRNASec purified on Sephacryl S-200 was phosphorylated with [γ-32P]ATP by tRNA kinase. The product [32P][3H]phosphoser-tRNA was purified on Sephacryl S-200 and hydrolyzed with ribonuclease T2. The chromatogram of this hydrolyzate on DEAE-cellulose in 7M urea buffer showed four peaks. The first peak of the pass-through fraction was seryl-adenosine liberated from the 3′-terminal of the tRNA. The second peak, eluted before the third peak containing inorganic phosphate, was phosphoseryl-adenosine. The major compound in the fourth peak was pGp. As a control experiment, non-acylated tRNASec was used as a substrate of phosphorylation and the product was analyzed. The chromatogram of the digest with ribonuclease T2 showed no peak of phosphoseryl-adenosine, but a peak of pGp was seen with the peak of inorganic phosphate. Thus, the major product in the presence of tRNA kinase was pGp, and a small but significant proportion of the radioactivity was found as phosphoserine in the presence of seryl residue on the 3′-CCA terminal of tRNASec. These results indicated that tRNA kinase phosphorylates not only Ser-tRNA to phosphoser-tRNA but also Gp of the 5′-termini of tRNA to pGp. This study gives a new role to mammalian tRNA kinase.
Similar content being viewed by others
References
Hatfield D & Portugal F (1970) Proc. Natl. Acad. Sci. USA 67: 1200–1206
Maenpaa PH & Bernfield MR (1970) Proc. Natl. Acad. Sci. USA 67: 688–695
Sharp SJ & Stewart TS (1977) Nucleic Acids Res. 4: 2123–2136
Mizutani T & Hashimoto A (1984) FEBS Lett. 169: 319–322
Lee BJ, Worland PJ, Davis JN, Stadtman, TC & Hatfield DJ (1989) J. Biol. Chem. 264: 9724–9727
Mizutani T, Kurata H, & Yamada K (1991) FEBS Lett. 289: 59–63
Mizutani T, Kurata H, Yamada K & Totsuka T (1992) Biochem. J. 284: 827–834
Mizutani T & Tachibana Y (1986) FEBS Lett. 207: 162–166
Mizutani T & Hitaka T (1988) FEBS Lett. 232: 243–248
Mizutani T, Maruyama N, Hitaka T & Sukenaga Y (1990) FEBS Lett. 269: 277
Hatfield D, Diamond A & Dudock B (1982) Proc. Natl. Acad. Sci. USA 79: 6215–6219
Lee BJ, de la Pena P, Tobian J, Zasloff M & Hatfield D (1987) Proc. Natl. Acad. Sci. USA 84: 6384–6388
Mizutani T, Kanbe K, Kimura Y, Tachibana Y & Hitaka T (1988) Chem. Pharm. Bull. 36: 824–827
Wu XQ & Gross HJ (1994) EMBO J. 13: 241–248
Hatfield D (ed) (2001) ‘Selenium: Its Molecular Biology and Role in Human Health’, Kluwer Academic Publishers, Boston, USA
Diamond A, Dudock B & Hatfield D (1981) Cell 25: 497–506
Amberg R, Urban C, Reuner B, Schaff P, Pomerantz S, McCloskey A & Gross HJ (1993) Nucleic Acids Res. 21: 5583–5588
McBride O, Rajagppalan M & Hatfield D (1987) J. Biol. Chem. 262: 11163–11166
Chambers I, Frampton J, Goldfarb P, Affala N, McBain W & Harrison P (1986) EMBO J. 5: 1221–1227
Sunde RA & Evenson JK (1987) J. Biol. Chem. 262: 933–937
Mizutani T, Narihara T & Hashimoto A (1984) Eur. J. Biochem. 143: 9–13
Mizutani T, Kanaya K & Tanabe K (1999) BioFactors 9: 27–36
Yamada K, Mizutani T, Ejiri S & Totsuka T (1994) FEBS Lett. 347: 137–142
Mizutani T, Tanabe K & Yamada K (1998) FEBS Lett. 429: 189–193
Fagegaltier D, Hubert N, Yamada K, Mizutani T, Carbon P & Krol A (2000) EMBO J. 19: 4796–4805
Ding F & Grabowski P (1999) RNA 5: 1561–1569
Gelpi C, Sontheimer EJ & Rodriguez-Sanchez JL (2000) Clin. Exp. Immunol. 121: 364–374
Berry MJ, Banu L, Chen Y, Mandel SJ, Kieffer JD, Harney JW & Larsen PR (1991) Nature 353: 273–276
Copeland P, Fletcher J, Carlson B, Hatfield D & Driscoll (2000) EMBO J. 19: 306–314
Atkins J & Gesteland R (2000) Nature 407: 463–464
Goto C, Osaka T & Mizutani T (2001) BioFactors 14: 25–35
Mizutani T & Fujiwara T (2000) Mol. Biol. Reports 27: 99–105
Korotkov KV, Kumaraswamy E, Zhou Y, Hatfield DL & Gladyshev VN (2001) J. Biol. Chem. 276: 15330–15336
Sturchler-Pierrat C, Hubert N, Totsuka T, Mizutani T, Carbon P & Krol A (1995) J. Biol. Chem. 270: 18570–18574
Amberg R & Mizutani T, Wu X & Gross HJ (1996) J. Mol. Biol. 263: 8–19
Sturchler C, Westhof E, Carbon P & Krol A (1993) Nucleic Acids Res. 21: 1073–1079
Bock A, Forchhammer K, Heider J & Baron C (1991) Trends Biochem. Sci. 16: 463–467
Mizutani T & Goto C (2000) FEBS Lett. 466: 359–362
Hubert N, Sturchler C, Westohof E, Carbon P & Krol A (1998) RNA 4: 1029–1033
Breitschopf K & Gross HJ (1994) EMBO J. 13: 3166–3169
Mizutani T, Kanaya K, Ikeda S, Fujiwara T, Yamada K & Totsuka T (1998) Mol. Biol. Reports 25: 211–216
Takemura S, Mizutani T & Miyazaki M (1968) J. Biochem. 64: 827–837
Gillam I, Millward S, Blew D, van Tigerstrom M, Wimmer E & Tener GM (1967) Biochemistry 6: 3043–3056
Kato N, Hoshino H & Harada F (1983) Biochem. Int. 7: 635–645
Mizutani T, Goto C & Totsuka T (2000) J. Health Sci. 46: 399–404
Jakubowski H (1997) Biochemistry 36: 11077–11085
Daimond AM, Montero-Puerner Y, Lee BJ & Hatfield D (1990) Nucleic Acids Res. 18: 6727
Low S, Harney W & Berry M (1995) J. Biol. Chem. 270: 21059–21664
Lee BJ, Rajagopalan M, Kim YS, You KH, Jacobson KB & Hatfield D (1990) Mol. Cell. Biol. 10: 1940–1949
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Mizutani, T., Osaka, T., Ito, Y. et al. pGp as the main product of bovine tRNA kinase. Mol Biol Rep 29, 293–300 (2002). https://doi.org/10.1023/A:1020423705963
Issue Date:
DOI: https://doi.org/10.1023/A:1020423705963