Proteins of viral and cellular origin bind to the Aleutian disease virus (ADV) DNA 3'-terminal hairpin: presentation of a scheme for encapsidation of ADV DNA

Abstract

We have observed the binding of viral and cellular proteins to the Aleutian disease virus (ADV) 3' terminus of replicative-form DNA. Gel retardation assays showed specific band shifts produced by whole-cell extracts from either ADV-infected or uninfected cells, as well as band reduction produced by ADV capsids. In all cases, binding was confined to the turnaround, T-shaped terminal form; no binding to the extended conformation of replicative-form DNA was detected. This indicates the importance of the T-shaped secondary structure in protein recognition. We have previously reported the binding of a 3'-terminal ADV DNA restriction fragment to the ADV capsid protein VP1 (K. Willwand and O.-R. Kaaden, Virology 166:52-57, 1988). Here we show that the region between nucleotides 14 and 102 on the ADV genome is required for binding. It is suggested that the VP1-DNA interaction mediates the binding of ADV DNA to empty viral capsids and that this is followed by displacement synthesis and packaging of ADV progeny DNA. A scheme for the possible mechanism of this process is presented.

The minor capsid protein VP1 of the autonomous parvovirus minute virus of mice is dispensable for encapsidation of progeny single-stranded DNA but is required for infectivity

“…These results might suggest that encapsidation of MVM viral DNA is dependent on VP1 and begins at the 3' end of the viral ssDNA after replication of the complementary strand is completed but as such would be in contrast to the finding of packaged MVM-defective DNA containing only the 5' end of viral DNA. Similar binding properties have also been reported for the VP1 of Aleutian disease virus (ADV) (66) and VP2 of bovine parvovirus (BPV) (32,39). (BPV produces an additional, larger 132 TULLIS ET AL.…”

Evidence for covalent attachment of the adeno-associated virus (AAV) rep protein to the ends of the AAV genome

“…Conceivably, the terminal protein promotes an interaction with capsid proteins and targets the associated DNA strand for packaging. In this regard, it is worth noting that the AAV packaging signal is located within the AAV terminal repeat (46,61) and that the capsid proteins of two other parvoviruses, Aleutian disease virus and bovine parvovirus (37,76), are capable of binding to their respective terminal repeats. Furthermore, in the case of LullI, it has been found that the removal of the terminal protein occurs concomitantly with viral packaging (49, 50).…”

The minute virus of mice capsid specifically recognizes the 3' hairpin structure of the viral replicative-form DNA: mapping of the binding site by hydroxyl radical footprinting

“…The binding was observed only for 3' ends in the hairpin conformation but not for extended forms (15,29). The ADV DNA had been shown to interact with only one of the two ADV capsid proteins, VP1 (28,29). Here we show that the 3' terminus of the MVM RF DNA in its hairpin conformation also binds to empty viral capsids and purified VP1.…”

“…Gel retardation assay. Band shift experiments were performed under the conditions described previously (29). Samples were analyzed by electrophoresis on 5% polyacrylamide gels ranging in length from 10 to 35 cm.…”

“…The RF DNAs of the autonomous parvoviruses Aleutian mink disease virus (ADV) and bovine parvovirus have been shown to interact with viral capsid proteins via their 3' termini. The binding was observed only for 3' ends in the hairpin conformation but not for extended forms (15,29). The ADV DNA had been shown to interact with only one of the two ADV capsid proteins, VP1 (28,29).…”