Adenovirus DNA-binding protein in cells infected with wild-type 5 adenovirus and two DNA-minus, temperature-sensitive mutants, H5ts125 and H5ts149.
Studies have been done to characterize further H5ts125, an adenovirus type 5 conditionally lethal, temperature-sensitive (ts) mutant defective in initiation of DNA synthesis and to investigate whether the single-strand-specific DNA-binding (72,000 molecular weight) protein is coded by the mutated viral gene. When H5ts125-infected cells were labeled with [35S]methionine at 32 degrees C and then incubated without isotope at 39.5 degrees C, the mutant's nonpermissive temperature, the 72,000 molecular weight polypeptide was progressively degraded. Immunofluorescence examination of cells infected with wild-type virus, H5ts125, and H5ts149 (a second, unique DNA-minus mutant) showed that immunologically reactive DNA-binding protein was barely detectable in H5ts125-infected cells at 39.5 degrees C, whereas this protein was present in wild-type- and H5TS149-infected cells, that the protein made at 32 degrees C in H5ts125-infected cells lost its ability to bind specific DNA-binding protein antibody when the infected cells were shifted to 39.5 degrees C, and that if H5ts125-infected cells were shifted from the restrictive temperature to 32 degrees C, even in the presence of cycloheximide to stop protein synthesis, immunologically reactive DNA-binding protein reappeared.
D000260:Adenoviruses, Human / Q000276:immunology / Q000378:metabolism*; D002352:Carrier Proteins / Q000276:immunology; D003513:Cycloheximide / Q000494:pharmacology; D004279:DNA, Viral / Q000096:biosynthesis* / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D009154:Mutation; D011485:Protein Binding; D013696:Temperature; D014764:Viral Proteins / Q000096:biosynthesis* / Q000276:immunology / Q000378:metabolism
D002352:Carrier Proteins; D004279:DNA, Viral; D014764:Viral Proteins; D003513:Cycloheximide