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Home / Equine Research Bank / J Bacteriol / Degradation of Deoxyribonucleic Acid and Alteration of Nucle...
Journal of bacteriology1963; 86(1); 138-146; doi: 10.1128/jb.86.1.138-146.1963

Degradation of Deoxyribonucleic Acid and Alteration of Nucleic Acid Metabolism in Suspension Cultures of L-M Cells Infected with Equine Abortion Virus.

Abstract: Randall, Charles C. (University of Mississippi School of Medicine, Jackson) and Barbara M. Walker. Degradation of deoxyribonucleic acid and alteration of nucleic acid metabolism in suspension cultures of L-M cells infected with equine abortion virus. J. Bacteriol. 86:138-146. 1963.-Metabolic alterations in log-phase suspension cultures infected with equine abortion virus (EAV) were determined in L-M cells simultaneously labeled or prelabeled with H(3)- or C(14)-thymidine. Although infection produced an early stimulation of the uptake of labeled thymidine (TdR) into the acid-soluble fraction of concurrently labeled cells, incorporation of the isotope into deoxyribonucleic acid (DNA) was progressively inhibited. The specific activity of infected-cell DNA was 48% of the control at 24 hr. The rate of incorporation of isotope from 12 to 24 hr was 43 and 13 counts per min per mug of DNA per hr for control and infected cultures, respectively. Owing to degradation of DNA, synthesis could not be accurately determined with the concurrently labeled cells. On the other hand, with prelabeled cells, quantitative isotopic methods could be used to determine the amount of DNA synthesized by measuring dilution of specific activity, even though infection triggered degradation of DNA into acid-soluble components. With this method, the DNA synthesized in infected cultures for 24 hr was approximately five times greater than the slight net increase determined by the diphenylamine reaction. The specific activity of infected-cell DNA decreased and then remained fixed after 24 hr, with 53% of the radioactivity appearing in the medium by 48 hr. No radioactive CO(2) was detected as a consequence of DNA degradation. Infected cells lost ribonucleic acid (RNA) as well as DNA; RNA and DNA were reduced by 64 and 50%, respectively, at 48 hr. The degradation of DNA was effectively inhibited by chelating agents in situ and is thought to be due to a deoxyribonuclease. Preliminary experiments with extracts of infected cells support this observation. The relationship of deoxyribonuclease to the synthesis of viral DNA remains to be determined.
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Publication Date: 1963-07-01 PubMed ID: 14051805PubMed Central: PMC278385DOI: 10.1128/jb.86.1.138-146.1963Google Scholar: Lookup
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  • Journal Article

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research by Charles C. Randall and Barbara M. Walker investigates how infection with equine abortion virus impacts deoxyribonucleic acids (DNA) and nucleic acid metabolism in suspension cultures of L-M cells.

Methods

The team utilized:

  • Suspension cultures of L-M cells in their log-phase, which were infected with the equine abortion virus (EAV).
  • These cells were either simultaneously labeled or prelabeled with H(3)- or C(14)-thymidine, isotopes used to study DNA replication and repair.
  • Quantitative isotopic methods with the prelabeled cells were used to determine the amount of DNA synthesized, despite infection-induced degradation of DNA.

Findings

In their results:

  • The researchers found that infection led to an initial stimulation of the uptake of labeled thymidine into the acid-soluble fraction of the concomitantly labeled cells.
  • However, as the infection progressed, the incorporation of the isotope into DNA was increasingly inhibited. Consequently, the specific activity of the DNA of the infected cells was reduced to 48% of the control at the 24-hour mark.
  • Despite the degradation of DNA due to the virus, it was found that the DNA synthesized in infected cultures 24 hours after infection was approximately five times greater than the slight net increase determined by the diphenylamine reaction.
  • Other results demonstrated that both ribonucleic acid (RNA) and DNA were reduced by 64% and 50% respectively in infected cells at 48 hours.

Conclusions

From their findings:

  • The researchers suggest that the degradation of DNA might be due to a deoxyribonuclease, a type of enzyme that catalyzes the hydrolytic cleavage of phosphodiester linkages in the DNA backbone.
  • This supposition is based on the fact that the degradation of DNA was effectively halted by chelating agents in situ. Preliminary experiments with extracts of infected cells support this observation.
  • However, the exact relationship between the deoxyribonuclease and the synthesis of viral DNA hasn’t been established and requires additional exploration.

This study contributes to understanding the strategies that viruses, like the equine abortion virus, may employ to interfere with host cellular processes, leading to a destabilization of nucleic acid metabolisms and DNA degradation in infected cells.

Cite This Article

APA
RANDALL CC, WALKER BM. (1963). Degradation of Deoxyribonucleic Acid and Alteration of Nucleic Acid Metabolism in Suspension Cultures of L-M Cells Infected with Equine Abortion Virus. J Bacteriol, 86(1), 138-146. https://doi.org/10.1128/jb.86.1.138-146.1963

Publication

Journal of bacteriology
ISSN: 0021-9193
NlmUniqueID: 2985120R
Country: United States
Language: English
Volume: 86
Issue: 1
Pages: 138-146

Researcher Affiliations

RANDALL, C C
    WALKER, B M

      MeSH Terms

      • Animals
      • DNA
      • DNA, Viral
      • Deoxyribonucleases
      • Female
      • Herpesviridae Infections
      • Herpesvirus 1, Equid
      • Horses
      • Metabolism
      • Pregnancy
      • RNA
      • RNA, Viral
      • Thymidine
      • Tissue Culture Techniques
      • Vertebrates
      • Viruses

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      Citations

      This article has been cited 5 times.
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