Reoviruses are prototypical members of the Reoviridae family of segmented dsRNA viruses that include important human (rotavirus) and veterinary (blue tongue disease) pathogens. The mechanisms by which 10 unique positive-sense viral RNAs are localized to viral factories for packaging into progeny core particles is poorly understood and will be addressed by the experiments in this proposal. The paths taken by reovirus positive-sense RNAs after they are transcribed will be examined by both in situ hybridization and immunostaining of bromouridine labeled viral positive-sense RNAs in infected cells. The mechanism for localization of viral RNAs to viral factories will be examined by microinjection studies of viral or control RNAs into reovirus infected cells, or transfected cells expressing reovirus RNA-binding proteins. If viral RNAs are specifically localized to viral factories, trans- and cis-acting factors involved in localization will be identified. Mechanisms for non-specific localization of RNA to factories will also be examined. Understanding the RNA localization process may lead to the development of new anti-viral strategies, and will aid in the development of a robust reverse genetics system for dsRNA viruses.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Postdoctoral Individual National Research Service Award (F32)
Project #
5F32AI056939-02
Application #
6794795
Study Section
Special Emphasis Panel (ZRG1-F08 (20))
Program Officer
Berard, Diana S
Project Start
2003-08-01
Project End
2005-07-31
Budget Start
2004-08-01
Budget End
2005-07-31
Support Year
2
Fiscal Year
2004
Total Cost
$48,928
Indirect Cost
Name
Harvard University
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
047006379
City
Boston
State
MA
Country
United States
Zip Code
02115
Miller, Cathy L; Arnold, Michelle M; Broering, Teresa J et al. (2010) Localization of mammalian orthoreovirus proteins to cytoplasmic factory-like structures via nonoverlapping regions of microNS. J Virol 84:867-82
Miller, Cathy L; Arnold, Michelle M; Broering, Teresa J et al. (2007) Virus-derived platforms for visualizing protein associations inside cells. Mol Cell Proteomics 6:1027-38
Broering, Teresa J; Arnold, Michelle M; Miller, Cathy L et al. (2005) Carboxyl-proximal regions of reovirus nonstructural protein muNS necessary and sufficient for forming factory-like inclusions. J Virol 79:6194-206
Miller, Cathy L; Parker, John S L; Dinoso, Jason B et al. (2004) Increased ubiquitination and other covariant phenotypes attributed to a strain- and temperature-dependent defect of reovirus core protein mu2. J Virol 78:10291-302