Skip to main content
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1980 Jan 25;8(2):225–234. doi: 10.1093/nar/8.2.225

Reinitiation of synthesis of small cytoplasmic RNA species K and L in isolated HeLa cell nuclei in vitro.

R Reichel, B J Benecke
PMCID: PMC327263  PMID: 7422543

Abstract

Isolated HeLa cell nuclei were used to synthesize low molecular weight RNA species in-vitro. The labelled RNA released from the nuclei during the incubation mainly consists of 5S RNA, pre-tRNA and small cytoplasmic RNA species K and L. All these low molecular weight RNA species are synthesized by RNA polymerase C (or III). The polyanion heparin was applied to study the reinitiation of these RNA molecules in-vitro. A comparison of the kinetics of RNA synthesis in the absence and in the presence of this inhibitor demonstrates a highly efficient in-vitro reinitiation of scRNA species K and L as well as 5S and pre-tRNA by RNA polymerase C. These results indicate a general competence of this enzyme to catalyze the de-novo formation of specific gene products in-vitro.

Full text

PDF
228

Images in this article

Selected References

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

  1. Benecke B. J., Penman S. A new class of small nuclear RNA molecules synthesized by a type I RNA polymerase in HeLa cells. Cell. 1977 Dec;12(4):939–946. doi: 10.1016/0092-8674(77)90158-1. [DOI] [PubMed] [Google Scholar]
  2. Benecke B. J., Penman S. Large species differences in the pattern of snPI RNA which can distinguish ape from human. J Cell Biol. 1979 Mar;80(3):778–783. doi: 10.1083/jcb.80.3.778. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Eliceiri G. L. Short-lived, small RNAs in the cytoplasm of HeLa cells. Cell. 1974 Sep;3(1):11–14. doi: 10.1016/0092-8674(74)90031-2. [DOI] [PubMed] [Google Scholar]
  4. Ferencz A., Seifart K. H. Comparative effect of heparin on RNA synthesis of isolated rat-liver nucleoli and purified RNA polymerase A. Eur J Biochem. 1975 May 6;53(2):605–613. doi: 10.1111/j.1432-1033.1975.tb04104.x. [DOI] [PubMed] [Google Scholar]
  5. Frederiksen S., Pedersen I. R., Hellung-Larsen P., Engberg J. Metabolic studies of small molecular weight nuclear RNA components in BHK-21 cells. Biochim Biophys Acta. 1974 Feb 27;340(1):64–76. doi: 10.1016/0005-2787(74)90174-9. [DOI] [PubMed] [Google Scholar]
  6. Goldstein L., Ko C. Electrophoretic characterization of shuttling and nonshuttling small nuclear RNAs. Cell. 1974 Aug;2(4):259–269. doi: 10.1016/0092-8674(74)90020-8. [DOI] [PubMed] [Google Scholar]
  7. Jensen E. G., Hellung-Larsen P., Frederiksen S. Synthesis of low molecular weight RNA components A, C and D by polymerase II in alpha-amanitin-resistant hamster cells. Nucleic Acids Res. 1979 Jan;6(1):321–330. doi: 10.1093/nar/6.1.321. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  9. Manley J. L., Sharp P. A., Gefter M. L. RNA synthesis in isolated nuclei: in vitro initiation of adenovirus 2 major late mRNA precursor. Proc Natl Acad Sci U S A. 1979 Jan;76(1):160–164. doi: 10.1073/pnas.76.1.160. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Marzluff W. F., Jr, Murphy E. C., Jr, Huang R. C. Transcription of ribonucleic acid in isolated mouse myeloma nuclei. Biochemistry. 1973 Aug 28;12(18):3440–3446. doi: 10.1021/bi00742a013. [DOI] [PubMed] [Google Scholar]
  11. Marzluff W. F., Jr, Murphy E. C., Jr, Huang R. C. Transcription of the genes for 5S ribosomal RNA and transfer RNA in isolated mouse myeloma cell nuclei. Biochemistry. 1974 Aug 27;13(18):3689–3696. doi: 10.1021/bi00715a011. [DOI] [PubMed] [Google Scholar]
  12. Penman S. RNA metabolism in the HeLa cell nucleus. J Mol Biol. 1966 May;17(1):117–130. doi: 10.1016/s0022-2836(66)80098-0. [DOI] [PubMed] [Google Scholar]
  13. Price R., Penman S. A distinct RNA polymerase activity, synthesizing 5-5 s, 5 s and 4 s RNA in nuclei from adenovirus 2-infected HeLa cells. J Mol Biol. 1972 Oct 14;70(3):435–450. doi: 10.1016/0022-2836(72)90551-7. [DOI] [PubMed] [Google Scholar]
  14. Udvardy A., Seifart K. H. Transcription of specific genes in isolated nuclei from HeLa cells in vitro. Eur J Biochem. 1976 Feb 16;62(2):353–363. doi: 10.1111/j.1432-1033.1976.tb10167.x. [DOI] [PubMed] [Google Scholar]
  15. Walter G., Zillig W., Palm P., Fuchs E. Initiation of DNA-dependent RNA synthesis and the effect of heparin on RNA polymerase. Eur J Biochem. 1967 Dec;3(2):194–201. doi: 10.1111/j.1432-1033.1967.tb19515.x. [DOI] [PubMed] [Google Scholar]
  16. Weil P. A., Luse D. S., Segall J., Roeder R. G. Selective and accurate initiation of transcription at the Ad2 major late promotor in a soluble system dependent on purified RNA polymerase II and DNA. Cell. 1979 Oct;18(2):469–484. doi: 10.1016/0092-8674(79)90065-5. [DOI] [PubMed] [Google Scholar]
  17. Weinberg R. A., Penman S. Small molecular weight monodisperse nuclear RNA. J Mol Biol. 1968 Dec;38(3):289–304. doi: 10.1016/0022-2836(68)90387-2. [DOI] [PubMed] [Google Scholar]
  18. Weinmann R., Raskas H. J., Roeder R. G. Role of DNA-dependent RNA polymerases II and III in transcription of the adenovirus genome late in productive infection. Proc Natl Acad Sci U S A. 1974 Sep;71(9):3426–3439. doi: 10.1073/pnas.71.9.3426. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Weinmann R., Roeder R. G. Role of DNA-dependent RNA polymerase 3 in the transcription of the tRNA and 5S RNA genes. Proc Natl Acad Sci U S A. 1974 May;71(5):1790–1794. doi: 10.1073/pnas.71.5.1790. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Zieve G., Benecke B. J., Penman S. Synthesis of two classes of small RNA species in vivo and in vitro. Biochemistry. 1977 Oct 4;16(20):4520–4525. doi: 10.1021/bi00639a029. [DOI] [PubMed] [Google Scholar]
  21. Zieve G., Penman S. Small RNA species of the HeLa cell: metabolism and subcellular localization. Cell. 1976 May;8(1):19–31. doi: 10.1016/0092-8674(76)90181-1. [DOI] [PubMed] [Google Scholar]
  22. Zylber E. A., Penman S. Products of RNA polymerases in HeLa cell nuclei. Proc Natl Acad Sci U S A. 1971 Nov;68(11):2861–2865. doi: 10.1073/pnas.68.11.2861. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES