Hepatitis B virus contains protein attached to the 5' terminus of its complete DNA strand.
Hepatitis B virus DNA contains a tightly bound protein which was not removed by healing to 60 degrees C with 2% SDS, 2% mercaptoethanol. The protein was indirectly demonstrated by the extraction of the DNA-protein complex with phenol before but not after its digestion with proteinase K. The DNA-protein complex had a lower buoyant density than protease-treated or free DNA; it was bound to glass fiber filters; it migrated at a slower rate in gel electrophoresis; and it could be radiolabeled by oxidative iodination. The binding site of the protein was mapped by extraction of restriction endonuclease digests with phenol and analysis of the digests for missing DNA fragments. The protein was localized to a site near the 5' end of the complete viral DNA strand. It remained attached to this strand after heating with SDS to 90 degrees C or treatment with 0.1 N NaOH, suggesting a covalent linkage. The 5' end of neither viral DNA strand could be phosphorylated in a reaction with polynucleotide kinase, consistent with attachment of protein to the 5' ends. The incomplete DNA strand, however, which is the strand elongated by the virion DNA polymerase reaction, did not contain a detectable amount of polypeptide as did the complete strand. The reasons for the apparent block of the 5' end of the incomplete DNA strand is thus not known. The protein bound covalently to HBV DNA could be involved in the replication of the complete viral DNA strand and/or endonucleolytic generation of linear unit-length DNA pieces from replicative intermediates, although its function and origin are not yet known.
D001665:Binding Sites; D055598:Chemical Phenomena; D002621:Chemistry; D004270:DNA, Circular / Q000378:metabolism*; D004279:DNA, Viral / Q000378:metabolism*; D006515:Hepatitis B virus / Q000032:analysis*; D006358:Hot Temperature; D010766:Phosphorylation; D011116:Polynucleotide 5'-Hydroxyl-Kinase / Q000378:metabolism; D014764:Viral Proteins / Q000378:metabolism*
D004270:DNA, Circular; D004279:DNA, Viral; D014764:Viral Proteins; D011116:Polynucleotide 5'-Hydroxyl-Kinase