Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1974 Feb;71(2):563–567. doi: 10.1073/pnas.71.2.563

Identification of Discrete Polyadenylate-containing RNA Components Transcribed from HeLa Cell Mitochondrial DNA*

Deanna Ojala 1, Giuseppe Attardi 1
PMCID: PMC388048  PMID: 4521823

Abstract

Polyacrylamide gel electrophoresis and sedimentation analysis under denaturing conditions of poly(A)-containing RNA from the polysome region of the sedimentation pattern of a HeLa-cell mitochondrial lysate has revealed the occurrence of a discrete RNA component, which sediments in the native state with a sedimentation constant of about 7 S. From the sedimentation behavior under native and denaturing conditions and the poly(A) content, a molecular weight of about 9 × 104 has been estimated for this component. RNA·DNA hybridization experiments have indicated that this component is coded for by the light strand of mitochondrial DNA. Evidence for the occurrence of a poly(A)-containing RNA component sedimenting at about 9 S and coded for by the heavy strand has also been obtained.

Keywords: actinomycin D, denaturation, poly(dT)-cellulose, polyacrylamide gel, separated strands, RNA·DNA hybridization

Full text

PDF
563

Selected References

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

  1. Aloni Y., Attardi G. Expression of the mitochondria genome in HeLa cells. IV. Titration of mitochondrial genes for 16 s, 12 s and 4 s RNA. J Mol Biol. 1971 Jan 28;55(2):271–276. doi: 10.1016/0022-2836(71)90197-5. [DOI] [PubMed] [Google Scholar]
  2. Aloni Y., Attardi G. Expression of the mitochondria genome in HeLa cells. IV. Titration of mitochondrial genes for 16 s, 12 s and 4 s RNA. J Mol Biol. 1971 Jan 28;55(2):271–276. doi: 10.1016/0022-2836(71)90197-5. [DOI] [PubMed] [Google Scholar]
  3. Aloni Y., Attardi G. Expression of the mitochondrial genome in HeLa cells. II. Evidence for complete transcription of mitochondrial DNA. J Mol Biol. 1971 Jan 28;55(2):251–267. doi: 10.1016/0022-2836(71)90195-1. [DOI] [PubMed] [Google Scholar]
  4. Aloni Y., Attardi G. Symmetrical in vivo transcription of mitochondrial DNA in HeLa cells. Proc Natl Acad Sci U S A. 1971 Aug;68(8):1757–1761. doi: 10.1073/pnas.68.8.1757. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Amaldi F., Attardi G. Partial sequence analysis of ribosomal RNA from HeLa cells. I. Oligonucleotide pattern of 28 s and 18 s RNA after pancreatic ribonuclease digestion. J Mol Biol. 1968 May 14;33(3):737–755. doi: 10.1016/0022-2836(68)90317-3. [DOI] [PubMed] [Google Scholar]
  6. Attardi B., Attardi G. Expression of the mitochondrial genome in HeLa cells. I. Properties of the discrete RNA components from the mitochondrial fraction. J Mol Biol. 1971 Jan 28;55(2):231–249. doi: 10.1016/0022-2836(71)90194-x. [DOI] [PubMed] [Google Scholar]
  7. Attardi G., Parnas H., Hwang M. I., Attardi B. Giant-size rapidly labeled nuclear ribonucleic acid and cytoplasmic messenger ribonucleic acid in immature duck erythrocytes. J Mol Biol. 1966 Sep;20(1):145–182. doi: 10.1016/0022-2836(66)90123-9. [DOI] [PubMed] [Google Scholar]
  8. Aviv H., Leder P. Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose. Proc Natl Acad Sci U S A. 1972 Jun;69(6):1408–1412. doi: 10.1073/pnas.69.6.1408. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Boedtker H. Dependence of the sedimentation coefficient on molecular weight of RNA after reaction with formaldehyde. J Mol Biol. 1968 Jul 14;35(1):61–70. doi: 10.1016/s0022-2836(68)80036-1. [DOI] [PubMed] [Google Scholar]
  10. Borst P., Grivell L. A. Mitochondrial ribosomes. FEBS Lett. 1971 Feb 19;13(2):73–88. doi: 10.1016/0014-5793(71)80204-1. [DOI] [PubMed] [Google Scholar]
  11. Darnell J. E., Philipson L., Wall R., Adesnik M. Polyadenylic acid sequences: role in conversion of nuclear RNA into messenger RNA. Science. 1971 Oct 29;174(4008):507–510. doi: 10.1126/science.174.4008.507. [DOI] [PubMed] [Google Scholar]
  12. Darnell J. E., Wall R., Tushinski R. J. An adenylic acid-rich sequence in messenger RNA of HeLa cells and its possible relationship to reiterated sites in DNA. Proc Natl Acad Sci U S A. 1971 Jun;68(6):1321–1325. doi: 10.1073/pnas.68.6.1321. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Dawid I. B., Chase J. W. Mitochondrial RNA in Xenopus laevis. II. Molecular weights and other physical properties of mitochondrial ribosomal and 4 s RNA. J Mol Biol. 1972 Jan 28;63(2):217–231. doi: 10.1016/0022-2836(72)90371-3. [DOI] [PubMed] [Google Scholar]
  14. Dubin D. T. The effect of actinomycin on the synthesis of mitochondrial RNA in hamster cells. Biochem Biophys Res Commun. 1967 Dec 15;29(5):655–660. doi: 10.1016/0006-291x(67)90266-5. [DOI] [PubMed] [Google Scholar]
  15. EIGNER J., BOEDTKER H., MICHAELS G. The thermal degradation of nucleic acids. Biochim Biophys Acta. 1961 Jul 22;51:165–168. doi: 10.1016/0006-3002(61)91028-9. [DOI] [PubMed] [Google Scholar]
  16. Edmonds M., Vaughan M. H., Jr, Nakazato H. Polyadenylic acid sequences in the heterogeneous nuclear RNA and rapidly-labeled polyribosomal RNA of HeLa cells: possible evidence for a precursor relationship. Proc Natl Acad Sci U S A. 1971 Jun;68(6):1336–1340. doi: 10.1073/pnas.68.6.1336. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Grivell L. A., Reijnders L., Borst P. The effect of temperature and ionic strength of the electrophoretic mobility of yeast mitochondrial RNA. Eur J Biochem. 1971 Mar 1;19(1):64–72. doi: 10.1111/j.1432-1033.1971.tb01288.x. [DOI] [PubMed] [Google Scholar]
  18. Jacob S. T., Schindler D. G., Morris H. P. Mitochondrial polyriboadenylate polymerase: relative lack of activity in hepatomas. Science. 1972 Nov 10;178(4061):639–640. doi: 10.1126/science.178.4061.639. [DOI] [PubMed] [Google Scholar]
  19. Jacob S. T., Schindler D. G. Polyriboadenylate polymerase solubilized from rat liver mitochondria. Biochem Biophys Res Commun. 1972 Jul 11;48(1):126–134. doi: 10.1016/0006-291x(72)90353-1. [DOI] [PubMed] [Google Scholar]
  20. Jeanteur P., Amaldi F., Attardi G. Partial sequence analysis of ribosomal RNA from HeLa cells. II. Evidence for sequences of non-ribosmal type in 45 and 32 s ribosomal RNA precursors. J Mol Biol. 1968 May 14;33(3):757–775. doi: 10.1016/0022-2836(68)90318-5. [DOI] [PubMed] [Google Scholar]
  21. Johnston R. E., Bose H. R. Correlation of messenger RNA function with adenylate-rich segments in the genomes of single-stranded RNA viruses. Proc Natl Acad Sci U S A. 1972 Jun;69(6):1514–1516. doi: 10.1073/pnas.69.6.1514. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Lai M. M., Duesberg P. H. Adenylic acid-rich sequence in RNAs of Rous sarcoma virus and Rauscher mouse leukaemia virus. Nature. 1972 Feb 18;235(5338):383–386. doi: 10.1038/235383c0. [DOI] [PubMed] [Google Scholar]
  23. Lederman M., Attardi G. Expression of the mitochondrial genome in HeLa cells. XVI. Electrophoretic properties of the products of in vivo and in vitro mitochondrial protein synthesis. J Mol Biol. 1973 Aug 5;78(2):275–283. doi: 10.1016/0022-2836(73)90116-2. [DOI] [PubMed] [Google Scholar]
  24. Lee S. Y., Mendecki J., Brawerman G. A polynucleotide segment rich in adenylic acid in the rapidly-labeled polyribosomal RNA component of mouse sarcoma 180 ascites cells. Proc Natl Acad Sci U S A. 1971 Jun;68(6):1331–1335. doi: 10.1073/pnas.68.6.1331. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Ojala D., Attardi G. Expression of the mitochondrial genome in HeLa cells. X. Properties of mitochondrial polysomes. J Mol Biol. 1972 Mar 28;65(2):273–289. doi: 10.1016/0022-2836(72)90282-3. [DOI] [PubMed] [Google Scholar]
  26. PERRY R. P. ROLE OF THE NUCLEOLUS IN RIBONUCLEIC ACID METABOLISM AND OTHER CELLULAR PROCESSES. Natl Cancer Inst Monogr. 1964 May;14:73–89. [PubMed] [Google Scholar]
  27. Penman S., Vesco C., Penman M. Localization and kinetics of formation of nuclear heterodisperse RNA, cytoplasmic heterodisperse RNA and polyribosome-associated messenger RNA in HeLa cells. J Mol Biol. 1968 May 28;34(1):49–60. doi: 10.1016/0022-2836(68)90234-9. [DOI] [PubMed] [Google Scholar]
  28. Perlman S., Abelson H. T., Penman S. Mitochondrial protein synthesis: RNA with the properties of Eukaryotic messenger RNA. Proc Natl Acad Sci U S A. 1973 Feb;70(2):350–353. doi: 10.1073/pnas.70.2.350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Philipson L., Wall R., Glickman G., Darnell J. E. Addition of polyadenylate sequences to virus-specific RNA during adenovirus replication. Proc Natl Acad Sci U S A. 1971 Nov;68(11):2806–2809. doi: 10.1073/pnas.68.11.2806. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. RICH A., DAVIES D. R., CRICK F. H., WATSON J. D. The molecular structure of polyadenylic acid. J Mol Biol. 1961 Feb;3:71–86. doi: 10.1016/s0022-2836(61)80009-0. [DOI] [PubMed] [Google Scholar]
  31. Robberson D., Aloni Y., Attardi G., Davidson N. Expression of the mitochondrial genome in HeLa cells. VI. Size determination of mitochondrial ribosomal RNA by electron microscopy. J Mol Biol. 1971 Sep 28;60(3):473–484. doi: 10.1016/0022-2836(71)90182-3. [DOI] [PubMed] [Google Scholar]
  32. SPIRIN A. S. The "temperature effect" and macromolecular structure of high-polymer ribonucleic acids of various origin. Biokhimiia. 1961 Nov-Dec;26:454–463. [PubMed] [Google Scholar]
  33. Tzagoloff A., Akai A. Assembly of the mitochondrial membrane system. 8. Properties of the products of mitochondrial protein synthesis in yeast. J Biol Chem. 1972 Oct 25;247(20):6517–6523. [PubMed] [Google Scholar]
  34. Weinberg R. A., Ben-Ishai Z., Newbold J. E. Poly A associated with SV40 messenger RNA. Nat New Biol. 1972 Jul 26;238(82):111–113. doi: 10.1038/newbio238111a0. [DOI] [PubMed] [Google Scholar]
  35. Wu M., Davidson N., Attardi G., Aloni Y. Expression of the mitochondrial genome in HeLa cells. XIV. The relative positions of the 4 S RNA genes and of the ribosomal RNA genes in mitochondrial DNA. J Mol Biol. 1972 Oct 28;71(1):81–93. doi: 10.1016/0022-2836(72)90402-0. [DOI] [PubMed] [Google Scholar]
  36. Zylber E., Vesco C., Penman S. Selective inhibition of the synthesis of mitochondria-associated RNA by ethidium bromide. J Mol Biol. 1969 Aug 28;44(1):195–204. doi: 10.1016/0022-2836(69)90414-8. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES