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. 1966 Jun;99(3):566–581. doi: 10.1042/bj0990566

Ribonucleic acid synthesis in yeast. The effect of cycloheximide on the synthesis of ribonucleic acid in Saccharomyces carlsbergensis

S R de Kloet 1
PMCID: PMC1265043  PMID: 5964958

Abstract

1. Cycloheximide causes the release of the control amino acids have over RNA synthesis in Saccharomyces carlsbergensis N.C.T.C. 74. 2. The antibiotic causes a gradual deceleration of RNA formation. After incubation for 60min. at 30° RNA synthesis usually proceeds at a rate only a few per cent of that of the untreated control. 3. In the presence of cycloheximide two types of RNA accumulate in the cell: soluble RNA and a high-molecular-weight RNA. The latter has a base composition intermediate between those of yeast DNA and yeast ribosomal RNA, and sediments in a sucrose gradient at a rate faster than that of the 23s ribosomal RNA component. 4. Yeast ribosomal RNA contains methylated bases. Judged from the incorporation of [Me-14C]methionine, the extent of methylation of ribosomal RNA is about 20% of that of the `soluble' RNA fraction. The high-molecular-weight RNA formed in the presence of cycloheximide is less methylated than normal RNA. In this case the sucrose-density-gradient sedimentation patterns of newly methylated and newly synthesized RNA do not coincide. 5. In the presence of cycloheximide, polysomal material accumulates, indicating that messenger RNA is formed. 6. The effect of the antibiotic on protein and RNA synthesis can be abolished by washing of the cells. The RNA that has accumulated during incubation of the cells with the antibiotic is not stable on removal of cycloheximide. 7. The results presented in this study are discussed in relation to the regulation of RNA formation in yeast.

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Selected References

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

  1. ARMSTRONG R. L., BOEZI J. A. STUDIES OF ESCHERICHIA COLI RIBONUCLEIC ACID-DEOXYRIBONUCLEIC ACID COMPLEX. Biochim Biophys Acta. 1965 May 11;103:60–69. doi: 10.1016/0005-2787(65)90541-1. [DOI] [PubMed] [Google Scholar]
  2. BENNETT L. L., Jr, SMITHERS D., WARD C. T. INHIBITION OF DNA SYNTHESIS IN MAMMALIAN CELLS BY ACTIDIONE. Biochim Biophys Acta. 1964 May 18;87:60–69. doi: 10.1016/0926-6550(64)90047-7. [DOI] [PubMed] [Google Scholar]
  3. BOLTON E. T., McCARTHY B. J. A general method for the isolation of RNA complementary to DNA. Proc Natl Acad Sci U S A. 1962 Aug;48:1390–1397. doi: 10.1073/pnas.48.8.1390. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. FAN D. P., HIGA A., LEVINTHAL C. MESSENGER RNA DECAY AND PROTECTION. J Mol Biol. 1964 Feb;8:210–222. doi: 10.1016/s0022-2836(64)80130-3. [DOI] [PubMed] [Google Scholar]
  5. FIALA E. S., DAVIS F. F. PREFERENTIAL INHIBITION OF SYNTHESIS AND METHYLATION OF RIBOSOMAL RNA IN NEUROSPORA CRASSA BY ACTIDIONE. Biochem Biophys Res Commun. 1965 Jan 4;18:115–118. doi: 10.1016/0006-291x(65)90892-2. [DOI] [PubMed] [Google Scholar]
  6. FLEISSNER E., BOREK E. A new enzyme of RNA synthesis: RNA methylase. Proc Natl Acad Sci U S A. 1962 Jul 15;48:1199–1203. doi: 10.1073/pnas.48.7.1199. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. GORDON J., BOMAN H. G., ISAKSSON L. A. IN VIVO INHIBITION OF RNA METHYLATION IN THE PRESENCE OF CHLORAMPHENICOL. J Mol Biol. 1964 Sep;9:831–833. doi: 10.1016/s0022-2836(64)80190-x. [DOI] [PubMed] [Google Scholar]
  8. HALVORSON H. Intracellular protein and nucleic acid turnover in resting yeast cells. Biochim Biophys Acta. 1958 Feb;27(2):255–266. doi: 10.1016/0006-3002(58)90332-9. [DOI] [PubMed] [Google Scholar]
  9. HENSHAW E. C. SUBUNITS OF RAT LIVER RIBOSOMES. J Mol Biol. 1964 Aug;9:610–612. doi: 10.1016/s0022-2836(64)80233-3. [DOI] [PubMed] [Google Scholar]
  10. HIATT H. H. A rapidly labeled RNA in rat liver nuclei. J Mol Biol. 1962 Aug;5:217–229. doi: 10.1016/s0022-2836(62)80085-0. [DOI] [PubMed] [Google Scholar]
  11. KERRIDGE D. The effect of actidione and other antifungal agents on nucleic acid and protein synthesis in Saccharomyces carlsbergensis. J Gen Microbiol. 1958 Dec;19(3):497–506. doi: 10.1099/00221287-19-3-497. [DOI] [PubMed] [Google Scholar]
  12. KITAZUME Y., YCAS M. THE CALCULATED COMPOSITION OF NEWLY SYNTHESIZED YEAST RIBONUCLEIC ACID. Biochim Biophys Acta. 1963 Nov 22;76:391–400. [PubMed] [Google Scholar]
  13. KURLAND C. G., MAALOE O. Regulation of ribosomal and transfer RNA synthesis. J Mol Biol. 1962 Mar;4:193–210. doi: 10.1016/s0022-2836(62)80051-5. [DOI] [PubMed] [Google Scholar]
  14. KURLAND C. G., NOMURA M., WATSON J. D. The physical properties of the chloromycetin particles. J Mol Biol. 1962 May;4:388–394. doi: 10.1016/s0022-2836(62)80019-9. [DOI] [PubMed] [Google Scholar]
  15. Kjellin-Straby K., Boman H. G. Studies on microbial RNA, 3. Formation of submethylated sRNA in Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1965 Jun;53(6):1346–1352. doi: 10.1073/pnas.53.6.1346. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. MIDGLEY J. E., McCARTHY B. J. The synthesis and kinetic behavior of deoxyribonucleic acid-like ribonucleic acid in bacteria. Biochim Biophys Acta. 1962 Nov 26;61:696–717. doi: 10.1016/0926-6550(62)90053-1. [DOI] [PubMed] [Google Scholar]
  17. MITCHISON J. M., GROSS P. R. SELECTIVE SYNTHESIS OF MESSENGER RNA IN A FISSION YEAST DURING A STEP-DOWN, AND ITS RELATION TO THE CELL CYCLE. BULK EXPERIMENTS. Exp Cell Res. 1965 Feb;37:259–277. doi: 10.1016/0014-4827(65)90175-8. [DOI] [PubMed] [Google Scholar]
  18. MONIER R., STEPHENSON M. L., ZAMECNIK P. C. The preparation and some properties of a low molecular weight ribonucleic acid from baker's yeast. Biochim Biophys Acta. 1960 Sep 9;43:1–8. doi: 10.1016/0006-3002(60)90399-1. [DOI] [PubMed] [Google Scholar]
  19. OSAWA S., TAKATA K., HOTTA Y. Nuclear and cytoplasmic ribonucleic acids of calf thymus. Biochim Biophys Acta. 1958 May;28(2):271–277. doi: 10.1016/0006-3002(58)90473-6. [DOI] [PubMed] [Google Scholar]
  20. SCHERRER K., LATHAM H., DARNELL J. E. Demonstration of an unstable RNA and of a precursor to ribosomal RNA in HeLa cells. Proc Natl Acad Sci U S A. 1963 Feb 15;49:240–248. doi: 10.1073/pnas.49.2.240. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. STENT G. S., BRENNER S. A genetic locus for the regulation of ribonucleic acid synthesis. Proc Natl Acad Sci U S A. 1961 Dec 15;47:2005–2014. doi: 10.1073/pnas.47.12.2005. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. TAKANAMI M., ZUBAY G. AN ESTIMATE OF THE SIZE OF THE RIBOSOMAL SITE FOR MESSENGER RNA BINDING. Proc Natl Acad Sci U S A. 1964 May;51:834–839. doi: 10.1073/pnas.51.5.834. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. TAMAOKI T., MUELLER G. C. Effect of puromycin on RNA synthesis in HeLa cells. Biochem Biophys Res Commun. 1963 May 28;11:404–410. doi: 10.1016/0006-291x(63)90131-1. [DOI] [PubMed] [Google Scholar]
  24. TISSIERES A., BOURGEOIS S., GROS F. Inhibition of RNA polymerase by RNA. J Mol Biol. 1963 Jul;7:100–103. doi: 10.1016/s0022-2836(63)80024-8. [DOI] [PubMed] [Google Scholar]
  25. WETTSTEIN F. O., NOLL H., PENMAN S. EFFECT OF CYCLOHEXIMIDE ON RIBOSOMAL AGGREGATES ENGAGED IN PROTEIN SYNTHESIS IN VITRO. Biochim Biophys Acta. 1964 Jul 22;87:525–528. doi: 10.1016/0926-6550(64)90131-8. [DOI] [PubMed] [Google Scholar]
  26. WILLIAMSON A. R., SCHWEET R. ROLE OF THE GENETIC MESSAGE IN POLYRIBOSOME FUNCTION. J Mol Biol. 1965 Feb;11:358–372. doi: 10.1016/s0022-2836(65)80063-8. [DOI] [PubMed] [Google Scholar]
  27. Whiffen A. J., Bohonos N., Emerson R. L. The Production of an Antifungal Antibiotic by Streptomyces griseus. J Bacteriol. 1946 Nov;52(5):610–611. doi: 10.1128/jb.52.5.610-611.1946. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. YCAS M., BRAWERMAN G. Interrelations between nucleic acid and protein biosynthesis in microorganisms. Arch Biochem Biophys. 1957 May;68(1):118–129. doi: 10.1016/0003-9861(57)90332-6. [DOI] [PubMed] [Google Scholar]
  29. ZAMENHOF S., CHARGAFF E., BRAWERMAN G. Dissymmetry in nucleotide sequence of desoxypentose nucleic acids. J Biol Chem. 1950 Nov;187(1):1–14. [PubMed] [Google Scholar]
  30. de KLOET S., van WERMESKERKEN R., KONINGSBERGER V. V. Studies on protein synthesis by protoplasts of Saccharomyces carlsbergensis. I. The effect of ribonuclease on protein synthesis. Biochim Biophys Acta. 1961 Feb 12;47:138–143. doi: 10.1016/0006-3002(61)90838-1. [DOI] [PubMed] [Google Scholar]
  31. de Kloet S. R. Accumulation of RNA with a DNA like base composition in Saccharomyces carlsbergensis in the presence of cycloheximide. Biochem Biophys Res Commun. 1965 May 18;19(5):582–586. doi: 10.1016/0006-291x(65)90378-5. [DOI] [PubMed] [Google Scholar]

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