Synthesis and Maturation of Ribosomal Ribonucleic Acids in Isolated HeLa Cell Nuclei. A Tracer Study on the Topology of the 45S Precursor of Ribosomal Ribonucleic Acids

Overview

Journal Biochem J

Specialty Biochemistry

Date 1974 Aug 1

PMID 4474879

Citations 4

Authors

A A HADJIOLOV

G I Milchev

Affiliations

Soon will be listed here.

Abstract

The synthesis and processing of RNA by isolated HeLa cell nuclei was studied at low ionic strength in the presence of alpha-amanitin. The RNA polymerase reaction, with endogenous template and enzyme, rapidly reaches a plateau dependent on the amount of nuclei. Evidence is presented that incorporation of [(3)H]UMP proceeds only in growing RNA chains, whereas initiation of new RNA chains is arrested. The product formed contains all the main components of the 45S pre-rRNA (precursor of rRNA) maturation pathway (45S, 32S and 20S pre-rRNA; 28S and 18S rRNA). Most of the labelled material is in the mature rRNA components and their immediate precursors, even at very short times of incubation (2min). Small, but definite, 5S and 4S RNA peaks are also observed. At shorter incubation times a substantial amount of [(3)H]UMP is incorporated into RNA molecules in the 24S and 10-16S zones. This RNA material is considered to represent the non-conserved segments of 45S pre-rRNA in the process of nucleolytic degradation. A model for the tracer study of the topology of 45S pre-rRNA, on arrest of rRNA initiation, is discussed. The experimental evidence obtained supports the following structure of 45S pre-rRNA: 5'-end-28S rRNA unit-18S rRNA unit-nonconserved segment-3'-end.

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References

Brown R, Haselkorn R . Synthesis and maturation of cytoplasmic ribosomal RNA in Euglena gracilis. J Mol Biol. 1971; 59(3):491-503. DOI: 10.1016/0022-2836(71)90312-3. View

Marzluff Jr W, Murphy Jr E, Huang R . Transcription of ribonucleic acid in isolated mouse myeloma nuclei. Biochemistry. 1973; 12(18):3440-6. DOI: 10.1021/bi00742a013. View

Willems M, Wagner E, Laing R, Penman S . Base composition of ribosomal RNA precursors in the HeLa cell nucleolus: further evidence of non-conservative processing. J Mol Biol. 1968; 32(2):211-20. DOI: 10.1016/0022-2836(68)90005-3. View

Fuchse , MILLETTE R, Zillig W, Walter G . Influence of salts on RNA synthesis by DNA-dependent RNA-polymerase from Escherichia coli. Eur J Biochem. 1967; 3(2):183-93. DOI: 10.1111/j.1432-1033.1967.tb19514.x. View

Moriyama Y, Choi Y, Busch H . Isolation and purification of a nuclear 4.4 S ribonucleic acid of the Novikoff hepatoma. J Biol Chem. 1970; 245(8):1970-7. View

Smith K, Arnstein H . RNA synthesis in isolated rabbit-liver nuclei. Eur J Biochem. 1972; 30(1):195-204. DOI: 10.1111/j.1432-1033.1972.tb02087.x. View

Hecht R, Birnstiel M . Integrity of the DNA template, a prerequisite for the faithful transcription of Xenopus rDNA in vitro. Eur J Biochem. 1972; 29(3):489-99. DOI: 10.1111/j.1432-1033.1972.tb02013.x. View

Reeder R, Roeder R . Ribosomal RNA synthesis in isolated nuclei. J Mol Biol. 1972; 67(3):433-41. DOI: 10.1016/0022-2836(72)90461-5. View

Grummt I, LINDIGKEIT R . Pre-ribosomal RNA synthesis in isolated rat-liver nucleoli. Eur J Biochem. 1973; 36(1):244-9. DOI: 10.1111/j.1432-1033.1973.tb02906.x. View

10.

CASTON J, Jones P . Synthesis and processing of high molecular weight RNA by nuclei isolated from embryos of Rana pipiens. J Mol Biol. 1972; 69(1):19-38. DOI: 10.1016/0022-2836(72)90021-6. View

11.

Weinberg R, Penman S . Processing of 45 s nucleolar RNA. J Mol Biol. 1970; 47(2):169-78. DOI: 10.1016/0022-2836(70)90337-2. View

12.

Burton K . A study of the conditions and mechanism of the diphenylamine reaction for the colorimetric estimation of deoxyribonucleic acid. Biochem J. 1956; 62(2):315-23. PMC: 1215910. DOI: 10.1042/bj0620315. View

13.

Younger L, Gelboin H . The electrophoretic distribution of RNA synthesized in vitro by isolated rat liver nuclei. Biochim Biophys Acta. 1970; 204(1):168-74. DOI: 10.1016/0005-2787(70)90499-5. View

14.

Siev M, Weinberg R, Penman S . The selective interruption of nucleolar RNA synthesis in HeLa cells by cordycepin. J Cell Biol. 1969; 41(2):510-20. PMC: 2107749. DOI: 10.1083/jcb.41.2.510. View

15.

HADJIOLOV A, DABEVA M, Mackedonski V . The action of alpha-amanitin in vivo on the synthesis and maturation of mouse liver ribonucleic acids. Biochem J. 1974; 138(3):321-34. PMC: 1166217. DOI: 10.1042/bj1380321a. View

16.

WIDNELL C, Tata J . A procedure for the isolation of enzymically active rat-liver nuclei. Biochem J. 1964; 92(2):313-7. PMC: 1205996. DOI: 10.1042/bj0920313. View

17.

Wellauer P, Dawid I . Secondary structure maps of RNA: processing of HeLa ribosomal RNA. Proc Natl Acad Sci U S A. 1973; 70(10):2827-31. PMC: 427118. DOI: 10.1073/pnas.70.10.2827. View

18.

Butterworth P, Cox R, Chesterton C . Transcription of mammalian chromatin by mammalian DNA-dependent RNA polymerases. Eur J Biochem. 1971; 23(2):229-41. DOI: 10.1111/j.1432-1033.1971.tb01613.x. View

19.

Sporn M . 2'-O-methylation of adenosine, guanosine, uridine, and cytidine in RNA of isolated rat liver nuclei. Proc Natl Acad Sci U S A. 1972; 69(7):1716-9. PMC: 426785. DOI: 10.1073/pnas.69.7.1716. View

20.

SMUCKLER E, HADJIOLOV A . Inhibition of hepatic deoxyribonucleic acid-dependent ribonucleic acid polymerases by the exotoxin of Bacillus thuringiensis in comparison with the effects of -amanitin and cordycepin. Biochem J. 1972; 129(1):153-66. PMC: 1174051. DOI: 10.1042/bj1290153. View