Complementation analysis of the combined Condit/Dales collection of vaccinia virus temperature-sensitive mutants has been reported (Lackner, C.A., D'Costa, S.M., Buck, C., Condit, R.C., 2003. Complementation analysis of the Dales collection of vaccinia virus temperature-sensitive mutants. Virology 305, 240-259), however not all complementation groups have previously been assigned to single genes on the viral genome. We have used marker rescue to map at least one representative of each complementation group to a unique viral gene. The final combined collection contains 124 temperature-sensitive mutants affecting 38 viral genes, plus five double mutants.

Background: Vaccinia protein H5 is a multifunctional protein involved in several aspects of viral replication. Results: H5 is a nucleic-acid binding, elongated rod-like tetramer with an instrinsically disordered N terminus and an ␣-helical C terminus. Conclusion: H5 functions as a hub protein in vaccinia virus replication linking otherwise unique viral processes. Significance: Understanding the mechanism of H5 function in vaccinia virus replication is crucial for studying vaccinia biology.