Infection of nondividing cells by Rous sarcoma virus

Abstract

A direct comparison demonstrates that Rous sarcoma virus is capable of infecting aphidicolin-arrested cells 10-fold more efficiently than murine leukemia virus but less efficiently than human immunodeficiency virus. The efficiency of infection of nondividing cells by the three viruses correlates with the respective ability of each viral DNA to enter the nucleus.

FIG. 1

Cell cycle analysis of HT1080 cells treated with aphidicolin. Cells were treated with aphidicolin for 24 h, washed, and maintained in the absence (−) or presence (+) of aphidicolin. The histograms show the results of fluorescence-activated cell sorter analyses after the cells had been stained with propidium iodide at the indicated times.

FIG. 2

Infectivity of HIV, RSV, and MLV on dividing and nondividing cells. Infections were carried out in aphidicolin-arrested cells that were released from or maintained in aphidicolin after infection. (A) Representative experiment of HT1080 cells infected with equivalent titers of HIV, RSV, or MLV and stained by X-Gal at 48 h postinfection. (B) Viral titers are expressed as LacZ infectious units per milliliter on HT1080 cells maintained in the absence (−) or presence (+) of aphidicolin. (C) Viral titers on nondividing cells are expressed as a percentage of titers obtained on dividing cells.

FIG. 2

Infectivity of HIV, RSV, and MLV on dividing and nondividing cells. Infections were carried out in aphidicolin-arrested cells that were released from or maintained in aphidicolin after infection. (A) Representative experiment of HT1080 cells infected with equivalent titers of HIV, RSV, or MLV and stained by X-Gal at 48 h postinfection. (B) Viral titers are expressed as LacZ infectious units per milliliter on HT1080 cells maintained in the absence (−) or presence (+) of aphidicolin. (C) Viral titers on nondividing cells are expressed as a percentage of titers obtained on dividing cells.

FIG. 2

Infectivity of HIV, RSV, and MLV on dividing and nondividing cells. Infections were carried out in aphidicolin-arrested cells that were released from or maintained in aphidicolin after infection. (A) Representative experiment of HT1080 cells infected with equivalent titers of HIV, RSV, or MLV and stained by X-Gal at 48 h postinfection. (B) Viral titers are expressed as LacZ infectious units per milliliter on HT1080 cells maintained in the absence (−) or presence (+) of aphidicolin. (C) Viral titers on nondividing cells are expressed as a percentage of titers obtained on dividing cells.

FIG. 3

PCR analysis of Hirt extracts from infected and uninfected cells. Aphidicolin-arrested HT1080 cells were infected with the indicated virus and released from or maintained in aphidicolin after infection. Hirt DNA was harvested 24 h postinfection and subjected to PCR analysis (see the text for details). (A) PCR amplification of elongated plus-strand viral DNA. Undiluted (Und.) Hirt DNA or Hirt DNA diluted 10-fold (1/10) or 100-fold (1/100) was used as a template for the PCR. (B) The same sample dilutions were used for PCR amplification of 2-LTR circular DNA. (C) PCR amplification of mitochondrial DNA from undiluted Hirt extracts from uninfected or infected cells that were maintained in the absence (−) or presence (+) of aphidicolin. H.I., undiluted Hirt DNA from cells incubated with heat-inactivated virus; mock, undiluted Hirt DNA from uninfected cells.