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. 1993 Jun;61(6):2383–2389. doi: 10.1128/iai.61.6.2383-2389.1993

Synthesis of DNA, rRNA, and protein by Rickettsia prowazekii growing in untreated or gamma interferon-treated mouse L929 cells.

Q Gao 1, J Turco 1, H H Winkler 1
PMCID: PMC280859  PMID: 7684727

Abstract

The syntheses of DNA, rRNA, and protein by Rickettsia prowazekii growing in mouse fibroblastic L929 cells were measured at various times after the addition of gamma interferon (IFN-gamma) to correlate the inhibition of a site of macromolecular synthesis with the established IFN-gamma-induced inhibition of rickettsial growth. A method was developed to measure the syntheses of DNA, rRNA, and protein by R. prowazekii during a 2-h pulse-labeling period while the rickettsiae were growing within cultured host cells that had intact macromolecular synthesis. This method involved incubation of the rickettsia-infected cells with a radioactive precursor (H3 32PO4 or Tran35S-label), purification of the rickettsiae, purification of rickettsial nucleic acids, and analysis of rickettsial nucleic acids and proteins by electrophoresis and autoradiography. A key feature of the method involved the use of calculated specific activities from a densitometric analysis of gels and autoradiograms, a procedure that made the data independent of rickettsial recovery. Rickettsial DNA and rRNA syntheses were both inhibited 12 h after the addition of IFN-gamma to infected cultures, whereas the synthesis of rickettsial proteins was not inhibited at this time. In contrast, at 20 h after the addition of IFN-gamma, rickettsial DNA, rRNA, and protein syntheses were all inhibited.

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Selected References

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