Interaction of normal and unusually modified microbial DNA with cultured mammalian cells. Breakdown and reincorporation vs. uptake of polymerized DNA.
The uptake of radioactively labeled bacterial and phage DNA and the incorporation of acid-soluble DNAase I digests of these DNAs by cultures of human foreskin and 3T3 cells were studied. The presence of large amounts of unusually modified pyrimidine residues in donor phage DNAs allowed radioactive donor DNA in the nuclei of DNA-treated cells to be distinguished from host DNA labeled with breakdown products derived from donor DNA. This distinction could be made because it was found that radioactively labeled 5-methylcytosine residues in predigested XP-12 DNA and glucosylated 5-hydroxymethylcytosine residues in predigested T4 DNA could not be incorporated in an unaltered form into animal cell DNA. The results obtained from the study of uptake of these DNAs suggest that approx. 4--40 ng of phage DNA per 10(6) cells was transported to the nuclei of DEAE-dextran-pretreated cells during 3 days of incubation in medium after treatment with the DNA. However, interpretation of the results is complicated by the finding of considerable amounts of donor DNA binding to and persisting at the cell surface, which might attach to nuclei during subcellular fractionation.
D001435:Bacteriophages; D001665:Binding Sites; D002412:Cations; D002462:Cell Membrane / Q000378:metabolism; D002467:Cell Nucleus / Q000378:metabolism; D002478:Cells, Cultured / Q000378:metabolism*; D004269:DNA, Bacterial / Q000378:metabolism*; D004279:DNA, Viral / Q000378:metabolism*; D011741:Pyrimidine Nucleosides / Q000378:metabolism; D013329:Structure-Activity Relationship
D002412:Cations; D004269:DNA, Bacterial; D004279:DNA, Viral; D011741:Pyrimidine Nucleosides