Skip to main content
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1979 Jul;76(7):3271–3275. doi: 10.1073/pnas.76.7.3271

A factor in serum and amniotic fluid is a substrate for the tRNA-modifying enzyme tRNA-guanine transferase.

J R Katze, W R Farkas
PMCID: PMC383806  PMID: 291001

Abstract

Q factor, a substance found in animal serum that enables cultured mammalian cells (L-M) to produce tRNA containing queuine (the base of "nucleoside Q", queuosine), has been purified to homogeneity from bovine amniotic fluid. Q factor causes the appearance of Q-containing tRNAAsp in the L-M cells cultivated in serum-free medium, and this was used as an assay to monitor the purification of Q factor. Q factor is a competitive inhibitor of guanine for rabbit reticulocyte tRNA-guanine trnsferase, with a K1 of 4.5 x 10(-8) M. Q factor is inactivated in both the L-M cell and tRNA-guanine transferase assays by treatment with periodate or cyanogen bromide, both of which react with queuine. In L-M cells, nearly complete conversion of Q-free to Q-containing tRNAAsp is observed within 24 hr after addition of pure Q factor to the medium; actinomycin D, cycloheximide, and cycloleucine, inhibitors of RNA synthesis, protein synthesis, and nucleic acid methylation, respectively, do not inhibit this conversion. The product of the reaction, catalyzed by pure rabbit reticulocyte tRNA-guanine transferase, between Q factor and rabbit reticulocyte Q-free tRNAHis is chromatographyically indistinguishable from Q-containing tRNAHis.

Full text

PDF
3271

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  2. Caboche M., Bachellerie J. P. RNA methylation and control of eukaryotic RNA biosynthesis. Effects of cycloleucine, a specific inhibitor of methylation, on ribosomal RNA maturation. Eur J Biochem. 1977 Mar 15;74(1):19–29. doi: 10.1111/j.1432-1033.1977.tb11362.x. [DOI] [PubMed] [Google Scholar]
  3. DIXON M. The determination of enzyme inhibitor constants. Biochem J. 1953 Aug;55(1):170–171. doi: 10.1042/bj0550170. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Dubrul E. F., Farkas W. R. Partial purification and properties of the reticulocyte guanylating enzyme. Biochim Biophys Acta. 1976 Sep 6;442(3):379–390. doi: 10.1016/0005-2787(76)90312-9. [DOI] [PubMed] [Google Scholar]
  5. ENNIS H. L., LUBIN M. CYCLOHEXIMIDE: ASPECTS OF INHIBITION OF PROTEIN SYNTHESIS IN MAMMALIAN CELLS. Science. 1964 Dec 11;146(3650):1474–1476. doi: 10.1126/science.146.3650.1474. [DOI] [PubMed] [Google Scholar]
  6. Farkas W. R., Chernoff D. Identification of the minor guanylated tRNA of rabbit reticulocytes. Nucleic Acids Res. 1976 Oct;3(10):2521–2528. doi: 10.1093/nar/3.10.2521. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Farkas W. R., Singh R. D. Guanylation of transfer ribonucleic acid by a cell-free lysate of rabbit reticulocytes. J Biol Chem. 1973 Nov 25;248(22):7780–7785. [PubMed] [Google Scholar]
  8. Hankins W. D., Farkas W. R. Guanylation of transfer RNA by rabbit reticulocytes. Biochim Biophys Acta. 1970 Jul 16;213(1):77–89. doi: 10.1016/0005-2787(70)90009-2. [DOI] [PubMed] [Google Scholar]
  9. Howes N. K., Farkas W. R. Studies with a homogeneous enzyme from rabbit erythrocytes catalyzing the insertion of guanine into tRNA. J Biol Chem. 1978 Dec 25;253(24):9082–9087. [PubMed] [Google Scholar]
  10. Kasai H., Nakanishi K., Macfarlane R. D., Torgerson D. F., Ohashi Z., McCloskey J. A., Gross H. J., Nishimura S. Letter: The structure of Q* nucleoside isolated from rabbit liver transfer ribonucleic acid. J Am Chem Soc. 1976 Aug 4;98(16):5044–5046. doi: 10.1021/ja00432a071. [DOI] [PubMed] [Google Scholar]
  11. Kasai H., Oashi Z., Harada F., Nishimura S., Oppenheimer N. J., Crain P. F., Liehr J. G., von Minden D. L., McCloskey J. A. Structure of the modified nucleoside Q isolated from Escherichia coli transfer ribonucleic acid. 7-(4,5-cis-Dihydroxy-1-cyclopenten-3-ylaminomethyl)-7-deazaguanosine. Biochemistry. 1975 Sep 23;14(19):4198–4208. doi: 10.1021/bi00690a008. [DOI] [PubMed] [Google Scholar]
  12. Katze J. R. Alterations in SVT2 cell transfer RNAs in response to cell density and serum type. Biochim Biophys Acta. 1975 Mar 10;383(2):131–139. doi: 10.1016/0005-2787(75)90254-3. [DOI] [PubMed] [Google Scholar]
  13. Katze J. R. Q-factor: a serum component required for the appearance of nucleoside Q in tRNA in tissue culture. Biochem Biophys Res Commun. 1978 Sep 29;84(2):527–535. doi: 10.1016/0006-291x(78)90201-2. [DOI] [PubMed] [Google Scholar]
  14. Katze J. R. Relation of cell type and cell density in tissue culture to the isoaccepting spectra of the nucleoside Q containing tRNAs: tRNATyr, tRNAHis, tRNAAsn and tRNAAsp. Nucleic Acids Res. 1978 Jul;5(7):2513–2524. doi: 10.1093/nar/5.7.2513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. McKinnon R. D., Wosnick M. A., White B. N. The role of the guanine insertion enzyme in O-biosynthesis in Drosophila melanogaster. Nucleic Acids Res. 1978 Dec;5(12):4865–4876. doi: 10.1093/nar/5.12.4865. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Noguchi S., Yamaizumi Z., Ohgi T., Goto T., Nishimura Y., Hirota Y., Nishimura S. Isolation of Q nucleoside precursor present in tRNA of an E. coli mutant and its characterization as 7-(cyano)-7-deazaguanosine. Nucleic Acids Res. 1978 Nov;5(11):4215–4223. doi: 10.1093/nar/5.11.4215. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Okada N., Noguchi S., Nishimura S., Ohgi T., Goto T., Crain P. F., McCloskey J. A. Structure determination of a nucleoside Q precursor isolated from E. coli tRNA: 7-(aminomethyl)-7-deazaguanosine. Nucleic Acids Res. 1978 Jul;5(7):2289–2296. doi: 10.1093/nar/5.7.2289. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Okada N., Shindo-Okada N., Sato S., Itoh Y. H., Oda K., Nishimura S. Detection of unique tRNA species in tumor tissues by Escherichia coli guanine insertion enzyme. Proc Natl Acad Sci U S A. 1978 Sep;75(9):4247–4251. doi: 10.1073/pnas.75.9.4247. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Okada N., Yasuda T., Nishimura S. Detection of nucleoside Q precursor in methyl-deficient E.coli tRNA. Nucleic Acids Res. 1977 Dec;4(12):4063–4075. doi: 10.1093/nar/4.12.4063. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Pearson R. L., Weiss J. F., Kelmers A. D. Improved separation of transfer RNA's on polychlorotrifuoroethylene-supported reversed-phase chromatography columns. Biochim Biophys Acta. 1971 Feb 11;228(3):770–774. doi: 10.1016/0005-2787(71)90748-9. [DOI] [PubMed] [Google Scholar]
  21. REICH E., FRANKLIN R. M., SHATKIN A. J., TATUMEL Action of actinomycin D on animal cells and viruses. Proc Natl Acad Sci U S A. 1962 Jul 15;48:1238–1245. doi: 10.1073/pnas.48.7.1238. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Roe B. A., Stankiewicz A. F., Chen C. Y. Chromatographic behavior of several mammalian tRNAs on acylated dihydroxyl-borate cellulose and Aminex A-28. Nucleic Acids Res. 1977 Jul;4(7):2191–2204. doi: 10.1093/nar/4.7.2191. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Roe B. A., Stankiewicz A. F., Rizi H. L., Weisz C., DiLauro M. N., Pike D., Chen C. Y., Chen E. Y. Comparison of rat liver and Walker 256 carcinosarcoma tRNAs. Nucleic Acids Res. 1979 Feb;6(2):673–688. doi: 10.1093/nar/6.2.673. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. White B. N., Tener G. M. Activity of a transfer RNA modifying enzyme during the development of Drosophila and its relationship to the su(s) locus. J Mol Biol. 1973 Mar 15;74(4):635–651. doi: 10.1016/0022-2836(73)90054-5. [DOI] [PubMed] [Google Scholar]
  25. Wilson T. G., Jacobson K. B. Isolation and characterization of pteridines from heads of Drosophila melanogaster by a modified thin-layer chromatography procedure. Biochem Genet. 1977 Apr;15(3-4):307–319. doi: 10.1007/BF00484462. [DOI] [PubMed] [Google Scholar]
  26. Yang W. K., Hellman A., Martin D. H., Hellman K. B., Novelli G. D. Isoaccepting transfer RNA's of L-M cells in culture and after tumor induction in C3H mice. Proc Natl Acad Sci U S A. 1969 Dec;64(4):1411–1418. doi: 10.1073/pnas.64.4.1411. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES