Is a fully wrapped SSB–DNA complex essential forEscherichia colisurvival?
| Autor: | Timothy M. Lohman, Elizabeth Weiland, Vincent M. Waldman, Alexander G. Kozlov |
|---|---|
| Rok vydání: | 2016 |
| Předmět: |
DNA
Bacterial 0301 basic medicine DNA Single-Stranded Plasma protein binding Biology medicine.disease_cause 03 medical and health sciences chemistry.chemical_compound stomatognathic system Oligosaccharide binding Escherichia coli Genetics medicine Binding site Binding Sites Microbial Viability Nucleic Acid Enzymes Oligonucleotide Escherichia coli Proteins Cooperative binding 3. Good health DNA-Binding Proteins stomatognathic diseases 030104 developmental biology Biochemistry chemistry Biophysics DNA DNA Damage Protein Binding Homotetramer |
| Zdroj: | Nucleic Acids Research |
| ISSN: | 1362-4962 0305-1048 |
| Popis: | Escherichia coli single-stranded DNA binding protein (SSB) is an essential homotetramer that binds ssDNA and recruits multiple proteins to their sites of action during genomic maintenance. Each SSB subunit contains an N-terminal globular oligonucleotide/oligosaccharide binding fold (OB-fold) and an intrinsically disordered C-terminal domain. SSB binds ssDNA in multiple modes in vitro, including the fully wrapped (SSB)65 and (SSB)56 modes, in which ssDNA contacts all four OB-folds, and the highly cooperative (SSB)35 mode, in which ssDNA contacts an average of only two OB-folds. These modes can both be populated under physiological conditions. While these different modes might be used for different functions, this has been difficult to assess. Here we used a dimeric SSB construct with two covalently linked OB-folds to disable ssDNA binding in two of the four OB-folds thus preventing formation of fully wrapped DNA complexes in vitro, although they retain a wild-type-like, salt-dependent shift in cooperative binding to ssDNA. These variants complement wild-type SSB in vivo indicating that a fully wrapped mode is not essential for function. These results do not preclude a normal function for a fully wrapped mode, but do indicate that E. coli tolerates some flexibility with regards to its SSB binding modes. |
| Databáze: | OpenAIRE |
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