It is increasingly appreciated that the recognition of specific RNA sequences by proteins is a fundamental aspect of a variety of normal cellular processes. Additional attention has been focussed on this problem because of the critical roles played by RNA-protein interactions in the life cycle of Human Immunodeficiency Virus, the cause of AIDS. The long term goal of this work is to contribute to a general understanding of the fundamental nature of RNA-protein interactions by understanding one particular model of such an interaction in detail. The coat protein of the bacterial virus MS2 is a genetic regulatory protein. It binds a specific RNA stem-loop (the translational operator) to repress translation of the viral replicase gene. Previous work in this laboratory has defined the RNA binding site through the isolation of coat protein mutants which either have reduced affinity for RNA, have increased affinity for RNA, or show altered RNA binding specificity. The main objective of the present proposal is to combine our separate knowledge of the structures of the coat protein RNA-binding site and its RNA ligand into a picture of how the two molecules interact. Two general approaches are proposed. First, since the mutant proteins must exert their effects by either removing contacts or adding new ones, the specific nature of these missing or added contacts can be probed by asking how mutant proteins differ from wild-type in their abilities to accommodate chemical modification or nucleotide substitution at specific sites in the RNA. Second, evolutionary variants of the MS2 virus utilize homologous coat proteins to bind divergent RNA structures. It is proposed to define the protein structural determinants of RNA binding specificity using a combination of genetic and biochemical methods.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM042901-05
Application #
2181727
Study Section
Microbial Physiology and Genetics Subcommittee 2 (MBC)
Project Start
1991-01-01
Project End
1997-12-31
Budget Start
1995-01-01
Budget End
1995-12-31
Support Year
5
Fiscal Year
1995
Total Cost
Indirect Cost
Name
University of New Mexico
Department
Anatomy/Cell Biology
Type
Schools of Medicine
DUNS #
829868723
City
Albuquerque
State
NM
Country
United States
Zip Code
87131
Lino, Christopher A; Caldeira, Jerri C; Peabody, David S (2017) Display of single-chain variable fragments on bacteriophage MS2 virus-like particles. J Nanobiotechnology 15:13
Frietze, Kathryn M; Peabody, David S; Chackerian, Bryce (2016) Engineering virus-like particles as vaccine platforms. Curr Opin Virol 18:44-9
O'Rourke, John P; Peabody, David S; Chackerian, Bryce (2015) Affinity selection of epitope-based vaccines using a bacteriophage virus-like particle platform. Curr Opin Virol 11:76-82
Caldeira, Jerri; Bustos, Jeremiah; Peabody, Julianne et al. (2015) Epitope-Specific Anti-hCG Vaccines on a Virus Like Particle Platform. PLoS One 10:e0141407
Caldeira, Jerri C; Peabody, David S (2011) Thermal stability of RNA phage virus-like particles displaying foreign peptides. J Nanobiotechnology 9:22
Ashley, Carlee E; Carnes, Eric C; Phillips, Genevieve K et al. (2011) Cell-specific delivery of diverse cargos by bacteriophage MS2 virus-like particles. ACS Nano 5:5729-45
Chackerian, Bryce; Caldeira, Jerri do Carmo; Peabody, Julianne et al. (2011) Peptide epitope identification by affinity selection on bacteriophage MS2 virus-like particles. J Mol Biol 409:225-37
Caldeira, Jerri do Carmo; Medford, Alexander; Kines, Rhonda C et al. (2010) Immunogenic display of diverse peptides, including a broadly cross-type neutralizing human papillomavirus L2 epitope, on virus-like particles of the RNA bacteriophage PP7. Vaccine 28:4384-93
Peabody, David S; Manifold-Wheeler, Brett; Medford, Alexander et al. (2008) Immunogenic display of diverse peptides on virus-like particles of RNA phage MS2. J Mol Biol 380:252-63
Ni, C Z; White, C A; Mitchell, R S et al. (1996) Crystal structure of the coat protein from the GA bacteriophage: model of the unassembled dimer. Protein Sci 5:2485-93

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