Properties of nucleoprotein complexes containing replicating polyoma DNA.
Short-lived nucleoprotein complexes (r-py complex) containing replicating polyoma DNA were isolated from infected cells after lysis with Triton X-100. The Triton lysing procedure of Green, Miller, and Hendler (1971) releases most complexes containing supercoiled viral DNA (py complex) from nuclei, but liberates only a portion of r-py complexes. r-py Complexes are associated more strongly with nuclear sites but can be extracted by prolonged incubation of nuclei in lysing solution. Complexes containing replicating polyoma DNA appear to be precursors to stable complexes containing supercoiled DNA. Sedimentation and buoyant density studies indicate that protein is bound to both r-py complexes and py complexes at a ratio of protein to DNA of about 1 to 2/1. Both types of complexes sediment as if the viral DNA is more compact than free DNA and both undergo major reversible configurational changes with increased salt concentration. Changes resulting from enzymatic and chemical treatment indicate that there may be two or more protein components in both r-py complex and py complex. One component is digested by Pronase and trypsin while another is resistant to the enzymes but released by deoxycholate. The abundance and similarity in chemical and physical properties of protein bound to all forms of polyoma DNA suggest that part of the protein molecules may serve in a structural capacity.
D000818:Animals; D000831:Animals, Newborn; D002458:Cell Fractionation; D002478:Cells, Cultured; D002499:Centrifugation, Density Gradient; D002586:Cesium; D002712:Chlorides; D004261:DNA Replication; D004279:DNA, Viral / Q000032:analysis* / Q000302:isolation & purification; D005557:Formaldehyde; D005976:Glutaral; D007668:Kidney; D051379:Mice; D009698:Nucleoproteins / Q000032:analysis* / Q000302:isolation & purification; D011120:Polyomavirus / Q000032:analysis*; D011402:Pronase; D012965:Sodium Chloride; D012972:Sodium Hydroxide; D013395:Sucrose; D013501:Surface-Active Agents; D014238:Trichloroacetic Acid; D014357:Trypsin
D002712:Chlorides; D004279:DNA, Viral; D009698:Nucleoproteins; D013501:Surface-Active Agents; D005557:Formaldehyde; D002586:Cesium; D012965:Sodium Chloride; D012972:Sodium Hydroxide; D013395:Sucrose; D014238:Trichloroacetic Acid; D014357:Trypsin; D011402:Pronase; D005976:Glutaral