1. The acidic C-terminus of vaccinia virus I3 single-strand binding protein promotes proper assembly of DNA-protein complexes.
- Author
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Harrison ML, Desaulniers MA, Noyce RS, and Evans DH
- Subjects
- Amino Acid Motifs, DNA Replication, DNA, Viral genetics, DNA-Binding Proteins chemistry, DNA-Binding Proteins genetics, Gene Expression Regulation, Viral, Humans, Vaccinia virus chemistry, Vaccinia virus genetics, Viral Proteins chemistry, Viral Proteins genetics, DNA, Viral metabolism, DNA-Binding Proteins metabolism, Vaccinia virology, Vaccinia virus metabolism, Viral Proteins metabolism
- Abstract
The vaccinia virus I3L gene encodes a single-stranded DNA binding protein (SSB) that is essential for virus DNA replication and is conserved in all Chordopoxviruses. The I3 protein contains a negatively charged C-terminal tail that is a common feature of SSBs. Such acidic tails are critical for SSB-dependent replication, recombination and repair. We cloned and purified variants of the I3 protein, along with a homolog from molluscum contagiosum virus, and tested how the acidic tail affected DNA-protein interactions. Deleting the C terminus of I3 enhanced the affinity for single-stranded DNA cellulose and gel shift analyses showed that it also altered the migration of I3-DNA complexes in agarose gels. Microinjecting an antibody against I3 into vaccinia-infected cells also selectively inhibited virus replication. We suggest that this domain promotes cooperative binding of I3 to DNA in a way that would maintain an open DNA configuration around a replication site., (Copyright © 2016 Elsevier Inc. All rights reserved.)
- Published
- 2016
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