Association of the histone-like protein HBsu with the nucleoid of Bacillus subtilis
Open Access
- 1 March 1997
- journal article
- Published by American Society for Microbiology in Journal of Bacteriology
- Vol. 179 (6), 2060-2064
- https://doi.org/10.1128/jb.179.6.2060-2064.1997
Abstract
To investigate the physiological role of the essential histone-like protein of Bacillus subtilis (HBsu) in the nucleoid structure, a fusion to the green fluorescent protein (GFP) of Aequorea victoria was constructed. This purified fusion protein, HBsuGFP, showed a threefold-reduced affinity to DNA compared to unmodified HBsu; however, in gel mobility shift experiments HBsuGFP DNA-binding was greatly enhanced in the presence of low HBsu concentrations. Additional production of HBsu also had a positive effect on the retarded growth of a B. subtilis strain, PK9C8, which expresses only hbs-gfp (encoding HBsuGFP). HBsu seemed to influence not only growth but also nucleoid structure, as monitored by DNA staining and fluorescence microscopy. Without HBsu production, strain PK9C8 showed a relaxed nucleoid structure associated with HBsuGFP. However, a highly compact nucleoid structure that coincides with the fluorescence of the fusion protein was visualized when HBsu synthesis was induced. This provides the first evidence for in vivo association of HBsu in DNA packaging and its consequence on cell growth.Keywords
This publication has 38 references indexed in Scilit:
- Structure and function of the Bacillus SpoIIE protein and its localization to sites of sporulation septum assemblyMolecular Microbiology, 1996
- Green Fluorescent Protein as a Marker for Gene ExpressionScience, 1994
- The DNA-binding protein HBsu is essential for normal growth and development in Bacillus subtilisBiochimie, 1992
- Requirement of integration host factor (IHF) for growth of Escherichia coli deficient in HU proteinGene, 1990
- Construction and characterization of the deletion mutant of hupA and hupB genes in Escherichia coliJournal of Molecular Biology, 1988
- Proteins from the prokaryotic nucleoid: primary and quaternary structure of the 15‐kD Escherichia coli DNA binding protein H‐NSMolecular Microbiology, 1988
- Primary structure of the hip gene of Escherichia coli and of its product, the β subunit of integration host factorJournal of Molecular Biology, 1985
- Isolation and Characterization of Small Heat-stable Acid-soluble DNA-binding Proteins from Bacillus subtilis NucleoidsMicrobiology, 1985
- “Western Blotting”: Electrophoretic transfer of proteins from sodium dodecyl sulfate-polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein AAnalytical Biochemistry, 1981
- A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye bindingAnalytical Biochemistry, 1976