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. 1976 May 1;69(2):229–240. doi: 10.1083/jcb.69.2.229

Action of dichlorobenzimidazole riboside on RNA synthesis in L-929 and HeLa cells

PMCID: PMC2109673  PMID: 1262389

Abstract

5,6-Dichloro-1-beta-D-ribofuranosylbenzimidazole (DRB) inhibits RNA synthesis in L-929 cells (mouse fibroblast line) and HeLa cells (human epitheloid carcinoma line) within 2 min of addition of the compound to the medium. By removing DRB from the medium, the inhibition is promptly and completely reversed after treatment of cells for as long as 1 h or even longer. The inhibitory effect of DRB on the overall rate of RNA synthesis is similar in L and HeLa cells and is markedly concentration- dependent in the low dose range (5-20 muM or 1.6-6.4 mug/ml), but not as higher concentrations of DRB. At a concentration of 12 muM, DRB has a highly selective inhibitory effect on the synthesis of nuclear heterogenous RNA in L cells. At higher concentrations, there is also inhibition of 45 S ribosomal precursor RNA synthesis, but at all concentrations the effect on heterogeneous RNA synthesis in L cells in considerably greater than that on preribosomal RNA synthesis. In HeLa cells, too, DRB has a selective effect on heterogeneous RNA synthesis, but quantitatively the selectivity of action is somewhat less pronounced. In both L and HeLa cells, the inhibition of synthesis of nuclear heterogeneous RNA is incomplete even at very high concentrations of DRB (150 muM). Thus, while DRB is a selective inhibitor of nuclear heterogeneous RNA synthesis, not all such RNA synthesis is sensitive to inhibition. It is proposed that messenger precursor RNA synthesis may largely be sensitive to inhibition by DRB. In short-term experiments, DRB has no effect on protein synthesis in L or HeLa cells. DRB has a slight to moderate inhibitory effect on uridine uptake into L cells and a moderate to marked effect on uptake of uridine into HeLa cells.

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Selected References

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

  1. ALLFREY V. G., MIRSKY A. E., OSAWA S. Protein synthesis in isolated cell nuclei. J Gen Physiol. 1957 Jan 20;40(3):451–490. doi: 10.1085/jgp.40.3.451. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Allfrey V. G., Mirsky A. E. SOME ASPECTS OF RIBONUCLEIC ACID SYNTHESIS IN ISOLATED CELL NUCLEI. Proc Natl Acad Sci U S A. 1957 Sep 15;43(9):821–826. doi: 10.1073/pnas.43.9.821. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bucknall R. A. The effects of substituted benzimidazoles on the growth of viruses and the nucleic acid metabolism of host cells. J Gen Virol. 1967 Jan;1(1):89–99. doi: 10.1099/0022-1317-1-1-89. [DOI] [PubMed] [Google Scholar]
  4. Cunningham D. D., Pardee A. B. Transport changes rapidly initiated by serum addition to "contact inhibited" 3T3 cells. Proc Natl Acad Sci U S A. 1969 Nov;64(3):1049–1056. doi: 10.1073/pnas.64.3.1049. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. EAGLE H. Amino acid metabolism in mammalian cell cultures. Science. 1959 Aug 21;130(3373):432–437. doi: 10.1126/science.130.3373.432. [DOI] [PubMed] [Google Scholar]
  6. Edström J. E., Egyházi E., Daneholt B., Lambert B., Ringborg U. Localization of newly synthesized low molecular weight RNA in polytene chromosomes from Chironomus tentans. Chromosoma. 1971;35(4):431–442. doi: 10.1007/BF02451449. [DOI] [PubMed] [Google Scholar]
  7. Egyházi E. A tentative initiation inhibitor of chromosomal heterogeneous RNA synthesis. J Mol Biol. 1974 Mar 25;84(1):173–183. doi: 10.1016/0022-2836(74)90220-4. [DOI] [PubMed] [Google Scholar]
  8. Egyházi E., Daneholt B., Edström J. E., Lambert B., Ringborg U. Intracellular distribution of low molecular weight RNA in Chironomus tentans. J Cell Biol. 1971 Jan;48(1):120–127. doi: 10.1083/jcb.48.1.120. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Fromageot H. P., Zinder N. D. Growth of bacteriophage f2 in E. coli treated with rifampicin. Proc Natl Acad Sci U S A. 1968 Sep;61(1):184–191. doi: 10.1073/pnas.61.1.184. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. GOMATOS P. J., TAMM I., DALES S., FRANKLIN R. M. Reovirus type 3: physical characteristics and interaction with L cells. Virology. 1962 Jul;17:441–454. doi: 10.1016/0042-6822(62)90139-3. [DOI] [PubMed] [Google Scholar]
  11. Granick D. Nucleolar necklaces in chick embryo fibroblast cells. I. Formation of necklaces by dichlororibobenzimidazole and other adenosine analogues that decrease RNA synthesis and degrade preribosomes. J Cell Biol. 1975 May;65(2):398–417. doi: 10.1083/jcb.65.2.398. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Granick D. Nucleolar necklaces in chick embryo fibroblast cells. II. Microscope observations of the effect of adenosine analogues on nucleolar necklace formation. J Cell Biol. 1975 May;65(2):418–427. doi: 10.1083/jcb.65.2.418. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  14. Mahy B. W., Hastie N. D., Armstrong S. J. Inhibition of influenza virus replication by -amanitin: mode of action. Proc Natl Acad Sci U S A. 1972 Jun;69(6):1421–1424. doi: 10.1073/pnas.69.6.1421. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Plagemann P. G., Richey D. P. Transport of nucleosides, nucleic acid bases, choline and glucose by animal cells in culture. Biochim Biophys Acta. 1974 Dec 16;344(3-4):263–305. doi: 10.1016/0304-4157(74)90010-0. [DOI] [PubMed] [Google Scholar]
  16. Ringborg U., Rydlander L. Nucleolar-derived ribonucleic acid in chromosomes, nuclear sap, and cytoplasm of Chironomus tentans salivary gland cells. J Cell Biol. 1971 Nov;51(21):355–368. doi: 10.1083/jcb.51.2.355. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. SIRLIN J. L., JACOB J. SEQUENTIAL AND REVERSIBLE INHIBITION OF SYNTHESIS OF RIBONUCLEIC ACID IN THE NUCLEOLUS AND CHROMOSOMES: EFFECT OF BENZAMIDE AND SUBSTITUTED BENZIMIDAZOLES ON DIPTERAN SALIVARY GLANDS. Nature. 1964 Nov 7;204:545–547. doi: 10.1038/204545a0. [DOI] [PubMed] [Google Scholar]
  18. Sehgal P. B., Tamm I., Vilcek J. Human interferon production: superinduction by 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole. Science. 1975 Oct 17;190(4211):282–284. doi: 10.1126/science.1179208. [DOI] [PubMed] [Google Scholar]
  19. Sehgal P. B., Tamm I., Vilcek J. Regulation of human interferon production. I. Superinduction by 5, 6-dichloro-1-beta-D-ribofuranosylbenzimidazole. Virology. 1976 Apr;70(2):532–541. doi: 10.1016/0042-6822(76)90294-4. [DOI] [PubMed] [Google Scholar]
  20. Sehgal P. B., Tamm I., Vilcek J. Regulation of human interferon production. II. Inhibition of interferon messenger RNA synthesis by 5, 6-dichloro-1-beta-D-ribofuranosylbenzimidazole. Virology. 1976 Apr;70(2):542–544. doi: 10.1016/0042-6822(76)90295-6. [DOI] [PubMed] [Google Scholar]
  21. Soeiro R., Vaughan M. H., Warner J. R., Darnell J. E., Jr The turnover of nuclear DNA-like RNA in HeLa cells. J Cell Biol. 1968 Oct;39(1):112–118. doi: 10.1083/jcb.39.1.112. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. TAMM I., FOLKERS K., SHUNK C. H., HORSFALL F. L., Jr Inhibition of influenza virus multiplication by N-glycosides of benzimidazoles-N. J Exp Med. 1954 Mar;99(3):227–250. doi: 10.1084/jem.99.3.227. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. TAMM I., FOLKERS K., SHUNK C. H. High inhibitory activity of certain halogenated ribofuranosylbenzimidazoles on influenza B virus multiplication. J Bacteriol. 1956 Jul;72(1):54–58. doi: 10.1128/jb.72.1.54-58.1956. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. TAMM I. Inhibition of influenza and mumps virus multiplication by 4,5,6- (or 5,6,7-) trichloro-1-beta-D-ribofuranosyl-benzimidazole. Science. 1954 Nov 19;120(3125):847–848. doi: 10.1126/science.120.3125.847. [DOI] [PubMed] [Google Scholar]
  25. TAMM I., NEMES M. M. Glycosides of chlorobenzimidazoles as inhibitors of poliovirus multiplication. Virology. 1957 Dec;4(3):483–498. doi: 10.1016/0042-6822(57)90081-8. [DOI] [PubMed] [Google Scholar]
  26. TAMM I., NEMES M. M., OSTERHOUT S. On the role of ribonucleic acid in animal virus synthesis. I. Studies with 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole. J Exp Med. 1960 Mar 1;111:339–349. doi: 10.1084/jem.111.3.339. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. TAMM I., OVERMAN J. R. Relationship between structure of benzimidazole derivatives and inhibitory activity on vaccinia virus multiplication. Virology. 1957 Feb;3(1):185–196. doi: 10.1016/0042-6822(57)90032-6. [DOI] [PubMed] [Google Scholar]
  28. TAMM I., TYRRELL D. A. Influenza virus multiplication in the chorioallantoic membrane in vitro: kinetic aspects of inhibition by 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole. J Exp Med. 1954 Dec 1;100(6):541–562. doi: 10.1084/jem.100.6.541. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Watanabe Y., Millward S., Graham A. F. Regulation of transcription of the Reovirus genome. J Mol Biol. 1968 Aug 28;36(1):107–123. doi: 10.1016/0022-2836(68)90223-4. [DOI] [PubMed] [Google Scholar]
  30. de Asua L. J., Rozengurt E., Dulbecco R. Kinetics of early changes in phosphate and uridine transport and cyclic AMP levels stimulated by serum in density-inhibited 3T3 cells. Proc Natl Acad Sci U S A. 1974 Jan;71(1):96–98. doi: 10.1073/pnas.71.1.96. [DOI] [PMC free article] [PubMed] [Google Scholar]

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