The central objective of the proposed research is to determine the molecular basis for protein-nucleic acid recognition which leads to the proper assembly and stability of simple plant and bacterial viruses.
The specific aims are: (i) to determine the secondary structures and conformations of nucleic acid and protein molecules in viruses, (ii) to identify the specific molecular subgroups which are responsible for intermolecular and intramolecular interactions in viruses, and (iii) to identify chemical factors, such as pH, ionic composition and the like, which control the proper assembly and disassembly of viruses. The above aims will be pursued primarily by laser-Raman spectroscopy of mature virions, of precursor particles isolated during plant virus or phage morphogenesis, and of purified capsid, procapsid, tail-spike and DNA-binding proteins. Viruses and viral components will also be investigated by electron microscopy and complementary spectroscopic methods. The interpretation of results on viruses will be aided by parallel studies of model nucleic acids and nucleoproteins. The data of Raman spectroscopy will be collected on a newly constructed and computerized laser Raman spectrometer employing both visible and near-ultraviolet lasers. The viruses targeted for initial study include representative filamentous viruses (fd, Ifl, Ike, Pfl, Xf, Pf3 and Cf), ds DNA viruses (P22 and T7), and RNA plant viruses (TMV and CCMV). The gp5-DNA complex of fd and corresponding complexes involving DNA binding proteins of other filamentous viruses will also be investigated.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI011855-11
Application #
3125040
Study Section
Biophysics and Biophysical Chemistry A Study Section (BBCA)
Project Start
1978-09-29
Project End
1988-08-31
Budget Start
1985-09-01
Budget End
1986-08-31
Support Year
11
Fiscal Year
1985
Total Cost
Indirect Cost
Name
Southeastern University of Massachusetts
Department
Type
Schools of Arts and Sciences
DUNS #
City
North Dartmouth
State
MA
Country
United States
Zip Code
Miura, T; Thomas Jr, G J (1995) Raman spectroscopy of proteins and their assemblies. Subcell Biochem 24:55-99
Prevelige Jr, P E; Thomas, D; Aubrey, K L et al. (1993) Subunit conformational changes accompanying bacteriophage P22 capsid maturation. Biochemistry 32:537-43
Li, H; Hanson, C; Fuchs, J A et al. (1993) Determination of the pKa values of active-center cysteines, cysteines-32 and -35, in Escherichia coli thioredoxin by Raman spectroscopy. Biochemistry 32:5800-8
Li, T; Bamford, D H; Bamford, J K et al. (1993) Structural studies of the enveloped dsRNA bacteriophage phi 6 of Pseudomonas syringae by Raman spectroscopy. I. The virion and its membrane envelope. J Mol Biol 230:461-72
Bamford, J K; Bamford, D H; Li, T et al. (1993) Structural studies of the enveloped dsRNA bacteriophage phi 6 of Pseudomonas syringae by Raman spectroscopy. II. Nucleocapsid structure and thermostability of the virion, nucleocapsid and polymerase complex. J Mol Biol 230:473-82
Li, T; Chen, Z; Johnson, J E et al. (1992) Conformations, interactions, and thermostabilities of RNA and proteins in bean pod mottle virus: investigation of solution and crystal structures by laser Raman spectroscopy. Biochemistry 31:6673-82
Aubrey, K L; Casjens, S R; Thomas Jr, G J (1992) Secondary structure and interactions of the packaged dsDNA genome of bacteriophage P22 investigated by Raman difference spectroscopy. Biochemistry 31:11835-42
Aubrey, K L; Thomas Jr, G J (1991) Raman spectroscopy of filamentous bacteriophage Ff (fd, M13, f1) incorporating specifically-deuterated alanine and tryptophan side chains. Assignments and structural interpretation. Biophys J 60:1337-49
Benevides, J M; Weiss, M A; Thomas Jr, G J (1991) Design of the helix-turn-helix motif: nonlocal effects of quaternary structure in DNA recognition investigated by laser Raman spectroscopy. Biochemistry 30:4381-8
Benevides, J M; Stow, P L; Ilag, L L et al. (1991) Differences in secondary structure between packaged and unpackaged single-stranded DNA of bacteriophage phi X174 determined by Raman spectroscopy: a model for phi X174 DNA packaging. Biochemistry 30:4855-63

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