Interaction of virally coded protein and a cell cycle-regulated cellular protein with the bovine parvovirus left terminus ori.
Replication of parvoviruses requires cis signals located in terminal palindromes that function as origins of replication in conjunction with trans-acting viral and cellular proteins. A gel retardation assay was used to identify proteins in crude nuclear extracts of bovine parvovirus (BPV)-infected bovine fetal lung cells that interact with the hairpinned left end (3' OH terminus of the viral minus strand in the flop conformation) of BPV. Three specific DNA-protein complexes formed. One complex was shown to involve a BPV structural protein(s) by inhibiting its formation when antiserum specific for these BPV proteins was used. By specific competition with serum containing antibodies against the BPV nonstructural proteins, a second complex was shown to involve a BPV nonstructural protein. A third complex contained protein of cellular origin and was also formed with extracts of uninfected bovine fetal lung cells. DNA competition assays suggest that the viral proteins do not bind to the right hairpin, which differs in sequence and secondary structure from the left terminus, or to a BPV terminus that lacks the first 52 nucleotides, preventing formation of the stem of the hairpin. The cellular protein is regulated in a cell cycle-dependent fashion, with its binding activity increased in uninfected, actively dividing cells compared with contact-inhibited cells. Since autonomous parvovirus replication requires an S-phase factor for progeny formation, the terminal binding protein demonstrated here is a candidate for this factor.
Metcalf
J B
JB
Biology Department, Virginia Polytechnic Institute and State University, Blacksburg 24061-0406.
Bates
R C
RC
Lederman
M
M
D000818:Animals; D002213:Capsid / Q000378:metabolism; D002417:Cattle; D002453:Cell Cycle; D004279:DNA, Viral / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism*; D009687:Nuclear Proteins / Q000378:metabolism*; D010321:Parvoviridae / Q000235:genetics* / Q000378:metabolism; D014758:Viral Core Proteins / Q000378:metabolism; D017361:Viral Nonstructural Proteins; D014764:Viral Proteins / Q000378:metabolism*; D014779:Virus Replication
D004279:DNA, Viral; D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; D014758:Viral Core Proteins; D017361:Viral Nonstructural Proteins; D014764:Viral Proteins