The Synthesis of Polyribonucleotides by Cytoplasmic Enzymes

Overview

Journal Biochem J

Specialty Biochemistry

Date 1966 May 1

PMID 5947148

Citations 8

Authors

J R Wykes

R M SMELLIE

Affiliations

Soon will be listed here.

Abstract

1. The possibility that the cell cytoplasm contains enzymes catalysing the biosynthesis of RNA was investigated in fractions obtained by differential centrifugation of homogenates of Landschutz ascites-tumour cells. 2. The microsomal fraction was shown to be most active in incorporating UMP residues from [alpha-(32)P]UTP into polyribonucleotide material. 3. The same fraction also incorporated [(3)H]CTP, [(3)H]ATP and [(3)H]GTP separately and independently of the presence of complementary ribonucleoside 5'-triphosphates. 4. The reaction was promoted by the addition of RNA and showed an absolute requirement for Mg(2+) ions. 5. Analysis of alkaline hydrolysates of the reaction products after the incorporation of [alpha-(32)P]UTP showed that most of the radioactivity was recovered in (2',3')-UMP residues irrespective of whether CTP, ATP and GTP were present in the reaction mixture. 6. Extraction of RNA from the reaction mixtures after the incorporation of [(3)H]ATP, [(3)H]GTP or [(3)H]CTP and analysis by sucrosedensity-gradient centrifugation showed no labelling of the ribosomal RNA. Radioactive material appeared between the 4s region and the meniscus of the sucrose gradient. In agreement with this observation, determinations of the chain length of the product showed that only short sequences of polynucleotides were synthesized. It is concluded that only homopolyribonucleotide synthesis is catalysed by the microsomal fractions and that there is little or no synthesis of RNA-like heteropolymers.

Citing Articles

Modification of ribonucleic Acid synthesis in nuclei isolated from normal and regenerating liver: some effects of salt and specific divalent cations.

Pogo A, Littau V, Allfrey V, Mirsky A Proc Natl Acad Sci U S A. 1967; 57(3):743-50.

PMID: 16591526 PMC: 335571. DOI: 10.1073/pnas.57.3.743.

Chain extension of ribonucleic acid by enzymes from rat liver cytoplasm.

Wilkie N, SMELLIE R Biochem J. 1968; 109(4):485-94.

PMID: 5683501 PMC: 1186934. DOI: 10.1042/bj1090485.

Ribonucleic acid polymerase catalyzing synthesis of double-stranded arbovirus ribonucleic acid.

Martin E, Sonnabend J J Virol. 1967; 1(1):97-109.

PMID: 5623960 PMC: 375509. DOI: 10.1128/JVI.1.1.97-109.1967.

Polyribonucleotide synthesis by subfractions of microsomes from rat liver.

Wilkie N, SMELLIE R Biochem J. 1968; 109(2):229-38.

PMID: 4971286 PMC: 1186779. DOI: 10.1042/bj1090229.

Transcribing complexes in cells infected by vesicular stomatitis virus and rabies virus.

Villarreal L, Holland J J Virol. 1974; 14(3):441-50.

PMID: 4369391 PMC: 355536. DOI: 10.1128/JVI.14.3.441-450.1974.

References

Britten R, Roberts R . High-Resolution Density Gradient Sedimentation Analysis. Science. 1960; 131(3392):32-3. DOI: 10.1126/science.131.3392.32. View

KLEMPERER H . The incorporation of AMP into oligoribonucleotides by an enzyme from rat liver. Biochem Biophys Res Commun. 1964; 15(3):269-72. DOI: 10.1016/0006-291x(64)90158-5. View

Baltimore D, Franklin R . A NEW RIBONUCLEIC ACID POLYMERASE APPEARING AFTER MENGOVIRUS INFECTION OF L-CELLS. J Biol Chem. 1963; 238:3395-400. View

Stevens A, Henry J . STUDIES ON THE RIBONUCLEIC ACID POLYMERASE FROM ESCHERICHIA COLI. I. PURIFICATION OF THE ENZYME AND STUDIES OF RIBONUCLEIC ACID FORMATION. J Biol Chem. 1964; 239:196-203. View

Nakamoto T, Fox C, Weiss S . ENZYMATIC SYNTHESIS OF RIBONUCLEIC ACID. I. PREPARATION OF RIBONUCLEIC ACID POLYMERASE FROM EXTRACTS OF MICROCOCCUS LYSODEIKTICUS. J Biol Chem. 1964; 239:167-74. View

Eason R, SMELLIE R . THE INTRACELLULAR SITE OF FORMATION OF ENCEPHALOMYOCARDITIS VIRUS RIBONUCLEIC ACID. J Biol Chem. 1965; 240:2580-6. View

Scherrer K, LATHAM H, Darnell J . Demonstration of an unstable RNA and of a precursor to ribosomal RNA in HeLa cells. Proc Natl Acad Sci U S A. 1963; 49:240-8. PMC: 299789. DOI: 10.1073/pnas.49.2.240. View

PERRY R, ERRERA M . The role of the nucleolus in ribonucleic acid-and protein synthesis. I. Incorporation of cytidine into normal and nucleolar inactivated HeLa cells. Biochim Biophys Acta. 1961; 49:47-57. DOI: 10.1016/0006-3002(61)90868-x. View

VENKATARAMAN P, Mahler H . Incorporation of adenylic acid into ribonucleic acid and synthetic polynucleotides. J Biol Chem. 1963; 238:1058-67. View

10.

Preiss J, Dieckmann M, Berg P . The enzymic synthesis of amino acyl derivatives of ribonucleic acid. IV. The formation of the 3'-hydroxyl terminal trinucleotide sequence of amino acid-acceptor ribonucleic acid. J Biol Chem. 1961; 236:1748-57. View

11.

BURDON R, SMELLIE R . The incorporation of uridine 5'-triphosphate into ribonucleic acid by enzyme fractions from Ehrlich ascites carcinoma cells. Biochim Biophys Acta. 1961; 51:153-62. DOI: 10.1016/0006-3002(61)91026-5. View

12.

Baltimore D, Eggers H, Franklin R, Tamm I . Poliovirus-induced RNA polymerase and the effects of virus-specific inhibitors on its production. Proc Natl Acad Sci U S A. 1963; 49:843-9. PMC: 300019. DOI: 10.1073/pnas.49.6.843. View

13.

Weissmann C, Borst P, BURDON R, Billeter M, Ochoa S . REPLICATION OF VIRAL RNA. IV. PROPERTIES OF RNA SYNTHETASE AND ENZYMATIC SYNTHESIS OF MS2 PHAGE RNA. Proc Natl Acad Sci U S A. 1964; 51:890-7. PMC: 300179. DOI: 10.1073/pnas.51.5.890. View

14.

Edmonds M, Abrams R . Nature of a polynucleotide required for polyribonucleotide formation from adenosine triphosphate with an enzyme from thymus nuclei. J Biol Chem. 1962; 237:2636-42. View

15.

Razzell W, Khorana H . Studies on polynucleotides. X. Enzymic degradation. Some properties and mode of action of spleen phosphodiesterase. J Biol Chem. 1961; 236:1144-9. View

16.

YANKOFSKY S, Spiegelman S . The identification of the ribosomal RNA cistron by sequence complementarity. II. Saturation of and competitive interaction at the RNA cistron. Proc Natl Acad Sci U S A. 1962; 48:1466-72. PMC: 220976. DOI: 10.1073/pnas.48.8.1466. View

17.

Razzell W, Khorana H . Studies on polynucleotides. III. Enzymic degradation; substrate specificity and properties of snake venom phosphodiesterase. J Biol Chem. 1959; 234(8):2105-13. View

18.

YANKOFSKY S, Spiegelman S . Distinct cistrons for the two ribosomal RNA components. Proc Natl Acad Sci U S A. 1963; 49:538-44. PMC: 299899. DOI: 10.1073/pnas.49.4.538. View

19.

Eason R, Cline M, SMELLIE R . RIBONUCLEIC ACIDS IN VIRUS-INFECTED AND UNINFECTED KREBS II ASCITES TUMOR CELLS. J Biol Chem. 1963; 238:3978-84. View

20.

Hecht L, Zamecnik P, Stephenson M, Scott J . Nucleoside tri-phosphates as precursors of ribonucleic acid end groups in a mammalian system. J Biol Chem. 1958; 233(4):954-63. View