| Autor: |
Roberts SM; Department of Microbiology, Immunology and Molecular Genetics, UT Health, San Antonio, TX 78229, USA., Aldis M; Department of Microbiology, Immunology and Molecular Genetics, UT Health, San Antonio, TX 78229, USA., Wright ET; Department of Biochemistry and Structural Biology, UT Health, San Antonio, TX 78229, USA., Gonzales CB; Department of Comprehensive Dentistry, UT Health, San Antonio, TX 78229, USA., Lai Z; Department of Molecular Medicine, UT Health, San Antonio, TX 78229, USA., Weintraub ST; Department of Biochemistry and Structural Biology, UT Health, San Antonio, TX 78229, USA., Hardies SC; Department of Biochemistry and Structural Biology, UT Health, San Antonio, TX 78229, USA., Serwer P; Department of Biochemistry and Structural Biology, UT Health, San Antonio, TX 78229, USA. |
| Abstrakt: |
Diversity of phage propagation, physical properties, and assembly promotes the use of phages in ecological studies and biomedicine. However, observed phage diversity is incomplete. Bacillus thuringiensis siphophage, 0105phi-7-2, first described here, significantly expands known phage diversity, as seen via in-plaque propagation, electron microscopy, whole genome sequencing/annotation, protein mass spectrometry, and native gel electrophoresis (AGE). Average plaque diameter vs. plaque-supporting agarose gel concentration plots reveal unusually steep conversion to large plaques as agarose concentration decreases below 0.2%. These large plaques sometimes have small satellites and are made larger by orthovanadate, an ATPase inhibitor. Phage head-host-cell binding is observed by electron microscopy. We hypothesize that this binding causes plaque size-increase via biofilm evolved, ATP stimulated ride-hitching on motile host cells by temporarily inactive phages. Phage 0105phi7-2 does not propagate in liquid culture. Genomic sequencing/annotation reveals history as temperate phage and distant similarity, in a virion-assembly gene cluster, to prototypical siphophage SPP1 of Bacillus subtilis . Phage 0105phi7-2 is distinct in (1) absence of head-assembly scaffolding via either separate protein or classically sized, head protein-embedded peptide, (2) producing partially condensed, head-expelled DNA, and (3) having a surface relatively poor in AGE-detected net negative charges, which is possibly correlated with observed low murine blood persistence. |
