Association of viral and plasmid DNA with the nuclear matrix during productive infection.
The association of simian virus 40 (SV40) DNA or plasmid DNA in subcellular fractions from either infected or transfected cells was examined. In lytically infected cells, approx. 25% of viral specific DNA during the infection cycle was retained in nuclei after washing with low ionic strength buffer and 1% Triton X-100. Viral replicating DNA found in the nuclear matrix was capable of performing limited DNA synthesis by the endogenous DNA polymerase in vitro. Viral DNA synthesized in vitro hybridized preferentially to SV40 Hind-III B and C fragments which are in proximity to the origin of replication. In plasmid-transfected COS-7 cells (SV40-transformed cells), the amount of plasmid DNA found in the nuclear matrix was related to its replication efficiency in cells. More than 80% of the plasmid DNA was tightly associated with subnuclear structures. Little or no plasmid DNA was found in the cytoplasmic fraction. The results suggest that, in extrachromosomal model systems, the association of DNA with nuclear matrix is important for the regulation of DNA replication.
Jones
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Department of Microbiology, University of Kansas, Lawrence 66045.
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D000818:Animals; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism*; D002472:Cell Transformation, Viral; D004261:DNA Replication; D004270:DNA, Circular / Q000378:metabolism; D004274:DNA, Recombinant / Q000378:metabolism; D004278:DNA, Superhelical / Q000378:metabolism; D004279:DNA, Viral / Q000096:biosynthesis / Q000378:metabolism*; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism; D009693:Nucleic Acid Hybridization; D010957:Plasmids; D013539:Simian virus 40 / Q000235:genetics*; D014162:Transfection
D004270:DNA, Circular; D004274:DNA, Recombinant; D004278:DNA, Superhelical; D004279:DNA, Viral; D004259:DNA-Directed DNA Polymerase