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. 1971 Aug;68(8):1757–1761. doi: 10.1073/pnas.68.8.1757

Symmetrical In Vivo Transcription of Mitochondrial DNA in HeLa Cells

Y Aloni 1, G Attardi 1
PMCID: PMC389287  PMID: 5288761

Abstract

RNA·DNA hybridization experiments utilizing separated strands of HeLa mitochondrial DNA and mit-RNA from HeLa cells exposed to short pulses of [5-3H]uridine have shown that the labeled RNA hybridizes with both the light (L) and the heavy (H) strand, though to a different relative extent depending upon the labeling time. Thus, hybridization of pulse-labeled RNA is about equal with the two strands when the pulse is very short (1-5 min), and becomes more and more predominant with the H strand with increasing pulse length. Pulse-labeled fast-sedimenting mit-RNA forms RNase-resistant double-stranded structures up to more than 5 μm long when self-annealed or annealed with an excess of unlabeled mit-RNA. These observations and the previous evidence of complete transcription of the H strand strongly suggest that mit-DNA is transcribed in HeLa cells symmetrically over a considerable portion of its length, with the transcript of the L strand being rapidly degraded or otherwise removed from the mitochondrial fraction.

Keywords: electron microscopy

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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 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]
  3. Attardi B., Attardi G. A membrane-associated RNA of cytoplasmic origin in HeLa cells. Proc Natl Acad Sci U S A. 1967 Sep;58(3):1051–1058. doi: 10.1073/pnas.58.3.1051. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. 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]
  5. Attardi B., Cravioto B., Attardi G. Membrane-bound ribosomes in HeLa cells. I. Their proportion to total cell ribosomes and their association with messenger RNA. J Mol Biol. 1969 Aug 28;44(1):47–70. doi: 10.1016/0022-2836(69)90404-5. [DOI] [PubMed] [Google Scholar]
  6. Bovre K., Szybalski W. Patterns of convergent and overlapping transcription within the b2 region of coliphage lambda. Virology. 1969 Aug;38(4):614–626. doi: 10.1016/0042-6822(69)90181-0. [DOI] [PubMed] [Google Scholar]
  7. Colby C., Duesberg P. H. Double-stranded RNA in vaccinia virus infected cells. Nature. 1969 Jun 7;222(5197):940–944. doi: 10.1038/222940a0. [DOI] [PubMed] [Google Scholar]
  8. Nass M. M., Buck C. A. Studies on mitochondrial tRNA from animal cells. II. Hybridization of aminoacyl-tRNA from rat liver mitochondria with heavy and light complementary strands of mitochondrial DNA. J Mol Biol. 1970 Dec 14;54(2):187–198. doi: 10.1016/0022-2836(70)90426-2. [DOI] [PubMed] [Google Scholar]
  9. Robberson D., Aloni Y., Attardi G. Electron microscopic visualization of mitochondrial RNA-DNA hybrids. J Mol Biol. 1971 Jan 28;55(2):267–270. doi: 10.1016/0022-2836(71)90196-3. [DOI] [PubMed] [Google Scholar]
  10. Saccone C., Aaij C., Borst P., Gadaleta M. N. Hybridization studies with RNA synthesized by isolated rat-liver mitochondria. Biochim Biophys Acta. 1970 Feb 18;199(2):373–381. doi: 10.1016/0005-2787(70)90080-8. [DOI] [PubMed] [Google Scholar]
  11. Tabak H. F., Borst P. Erroneous transcription of mitochondrial DNA by RNA polymerase from Escherichia coli. Biochim Biophys Acta. 1970 Oct 15;217(2):356–364. doi: 10.1016/0005-2787(70)90533-2. [DOI] [PubMed] [Google Scholar]
  12. Wolfe M. W., Amsterdam D. Plaquelike Clearings Induced by Antimicrobial Agents on Lawns of Pseudomonas aeruginosa. Infect Immun. 1970 Feb;1(2):190–194. doi: 10.1128/iai.1.2.190-194.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Yoshiike K. Studies on DNA from low-density particles of SV40. I. Heterogeneous defective virions produced by successive undiluted passages. Virology. 1968 Mar;34(3):391–401. doi: 10.1016/0042-6822(68)90059-7. [DOI] [PubMed] [Google Scholar]

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