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 viruses and gene regulatory complexes.
The specific aims are to determine, for native viruses and macromolecular assemblies, the following: (i) specific molecular subgroups which are responsible for protein-nucleic acid recognition, (ii) chemical and physical factors, such as pH, ionic composition, and temperature, which regulate assembly and disassembly processes, and (iii) secondary structures and conformations of nucleic acid and protein molecules at physiological conditions.
These aims shall be pursued primarily by the method of laser Raman spectroscopy, employing the powerful technique of optical multichannel analysis (OMA). Targeted for study are a number of viruses and nucleoproteins for which complementary genetic, biochemical and structural information is available or obtainable. Among these systems are filamentious bacteriophages, isometric viruses, and phage lambda cI and Cro repressors and their recognition operators. The interpretation of results on viruses and nucleoproteins will be aided by parallel studies of model DNA and protein molecules. The collection of Raman spectroscopic data by the OMA rapid acquisition technique will enable the investigation of both equilibrium structures and dynamic processes. Structures of viruses and nucleoproteins will be determined and compared in both the solution and crystal states. The approach is one which can test the applicability of X-ray determined crystal structures for aqueous nucleic acids, proteins and their biologically active assemblies. Since many of the results will be applicable to viruses which infect higher organisms, this research program has long range health-related benefits.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI011855-16
Application #
3125043
Study Section
Biophysical Chemistry Study Section (BBCB)
Project Start
1987-09-01
Project End
1993-08-31
Budget Start
1990-09-01
Budget End
1991-08-31
Support Year
16
Fiscal Year
1990
Total Cost
Indirect Cost
Name
University of Missouri Kansas City
Department
Type
Schools of Medicine
DUNS #
800772162
City
Kansas City
State
MO
Country
United States
Zip Code
64110
Miura, T; Thomas Jr, G J (1995) Raman spectroscopy of proteins and their assemblies. Subcell Biochem 24:55-99
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
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; 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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