Human parainfluenza virus type 3 (HPIV3) is an important respiratory pathogen of infants, young children, elderly and immunocomprimised individuals for which an effective vaccine or treatment is not currently available. The goals of this proposal are to gain a thorough understanding of the viral mechanisms of RNA transcription and replication, and to develop a system to analyze the assembly/release and attachment/entry steps of the HPIV3 replicative cycle. A minigenome reverse genetics system, recently developed for HPIV3, will be used to delineate the sequences involved in promoting HPIV3 RNA replication and transcription (specific aims 1 and 2).
The first aim i nvolves analysis of a large RNA region of questionable involvement in replication, with the goal of unequivocally defining the significance of this region in replication.
The second aim seeks to determine whether a particular sequence serves as part of the promoter for transcription, thereby distinguishing a promoter for transcription from a promoter for replication.
Specific aim 3 seeks to develop the established minigenome system for HPIV3, which allows analysis RNA replication and transcription, into a system in which virus-like particles can be produced. This will allow for functional analysis of the assembly/release and attachment/entry steps of the viral life cycle in a convenient, efficient manner. Establishing such a system will allow for future study of the protein-protein interactions involved in these processes, and could thereby provide target interactions for drug development.
| Gander, Jill R; Schwan, LeeAnne M; Hoffman, Michael A (2011) Analysis of nucleotides 13-96 of the human parainfluenza virus type 3 antigenomic promoter reveals positive- and negative-acting replication elements. Virology 419:90-6 |
| Hoffman, Michael A; Thorson, LeeAnne M; Vickman, John E et al. (2006) Roles of human parainfluenza virus type 3 bases 13 to 78 in replication and transcription: identification of an additional replication promoter element and evidence for internal transcription initiation. J Virol 80:5388-96 |
