Structure and Organization of Bacteriophage Pf3 Probed by Raman and Ultraviolet Resonance Raman Spectroscopy
| Autor: | Stacy A. Overman, George J. Thomas, Priya Bondre, Zai-Qing Wen |
|---|---|
| Rok vydání: | 2000 |
| Předmět: |
Base pair
Protein subunit Resonance Raman spectroscopy Deoxyribonucleosides Arginine Spectrum Analysis Raman Biochemistry chemistry.chemical_compound symbols.namesake Capsid Deuterium Oxide Base Pairing Protein secondary structure Virus Assembly Tryptophan Bacteriophage Pf1 Water Hydrogen Bonding Amides Solutions Crystallography Spectrometry Fluorescence chemistry Virion assembly DNA Viral symbols Capsid Proteins Spectrophotometry Ultraviolet Peptides Pseudomonas Phages Raman spectroscopy Thymine DNA |
| Zdroj: | Biochemistry. 40:449-458 |
| ISSN: | 1520-4995 0006-2960 |
| DOI: | 10.1021/bi0018887 |
| Popis: | The Pseudomonas bacteriophage Pf3 is a long and narrow filament consisting of a covalently closed DNA single strand of 5833 bases sheathed by approximately 2500 copies of a 44-residue subunit. Ultraviolet resonance Raman spectra excited at 257, 244, 238, and 229 nm and off-resonance Raman spectra excited at 514.5 nm are reported for Pf3 in both H2O and D2O solutions. The key Raman bands are assigned to specific protein and DNA groups of the native virion assembly. The results are compared with proposed assembly models and Raman spectra recently reported for the isomorphous (class II) Pseudomonas phage Pf1 and the morphologically distinct (class I) coliphage fd [Wen, Z. Q., Overman, S. A., and Thomas, G. J. , Jr. (1997) Biochemistry 36, 7810-7820; Wen, Z. Q., Armstrong, A., and Thomas, G. J., Jr. (1999) Biochemistry 38, 3148-3156]. Surprisingly, deoxynucleosides of the packaged DNA genome of Pf3 adopt the same conformation (C3'-endo/anti) found for DNA packaged in the class I fd virion rather than that (C2'-endo/anti) associated with DNA in the isomorphous Pf1 virion. However, DNA base stacking in Pf3, as judged by Raman hypochromic effects, differs significantly from that occurring in either Pf1 or fd. Thus, the single-stranded DNA genomes of Pf3, Pf1, and fd are all organized differently within their respective capsids, implying that local subunit-DNA interactions may be important in determining the structure specific to each native assembly. The present study confirms a completely alpha-helical secondary structure for the Pf3 subunit and an unusual indolyl ring environment for the subunit tryptophan residue (Trp-38). |
| Databáze: | OpenAIRE |
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