UvrB protein of Corynebacterium pseudotuberculosis complements the phenotype of knockout Escherichia coli and recognizes DNA damage caused by UV radiation but not 8-oxoguanine in vitro
| Autor: | Rafael Cançado de Faria, Bárbara Catarina Teodoro Castro, Débora de Oliveira Lopes, Vasco Azevedo, Carlos Renato Machado, Moacyr Comar Junior, Luciana Lara dos Santos, Bruna Franciele Faria |
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| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Guanine Ultraviolet Rays DNA repair DNA damage Corynebacterium pseudotuberculosis Pyrimidine dimer Biology medicine.disease_cause 03 medical and health sciences chemistry.chemical_compound Bacterial Proteins Escherichia coli Genetics medicine Amino Acid Sequence Cloning Molecular Sequence Homology Amino Acid Oligonucleotide General Medicine Molecular biology 030104 developmental biology chemistry Gene Knockdown Techniques DNA DNA Damage Nucleotide excision repair |
| Zdroj: | Gene. 639:34-43 |
| ISSN: | 0378-1119 |
| DOI: | 10.1016/j.gene.2017.09.068 |
| Popis: | In prokaryotic cells, the UvrB protein plays a central role in nucleotide excision repair, which is involved in the recognition of bulky DNA lesions generated by chemical or physical agents. The present investigation aimed to characterize the uvrB gene of Corynebacterium pseudotuberculosis (CpuvrB) and evaluate its involvement in the DNA repair system of this pathogenic organism. In computational analysis, the alignment of the UvrB protein sequences of Escherichia coli, Mycobacterium tuberculosis, Bacillus caldotenax and Corynebacterium pseudotuberculosis showed high similarity and the catalytic amino acid residues and functional domains are preserved. A CpUvrB model was constructed by comparative modeling and presented structural similarity with the UvrB of E. coli. Moreover, in molecular docking analysis CpUvrB showed favorable interaction with EcUvrA and revealed a preserved ATP incorporation site. Heterologous functional complementation assays using E. coli uvrB-deficient cells exposed to UV irradiation showed that the CpUvrB protein contributed to an increased survival rate in relation to those in the absence of CpUvrB. Damaged oligonucleotides containing thymine dimer or 8-oxoguanine lesion were synthesized and incubated with CpUvrB protein, which was able to recognize and excise UV irradiation damage but not 8-oxoguanine. These results suggest that CpUvrB is involved in repairing lesions derived from UV light and encodes a protein orthologous to EcUvrB. |
| Databáze: | OpenAIRE |
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