DNA binding properties and localization of paralogous SSB proteins from Streptomyces coelicolor (CROSBI ID 695327)
Prilog sa skupa u zborniku | sažetak izlaganja sa skupa | međunarodna recenzija
Podaci o odgovornosti
Tina Paradžik ; Vlatka Godinić Mikulčić ; Anja Kostelac ; Dušica Vujaklija
engleski
DNA binding properties and localization of paralogous SSB proteins from Streptomyces coelicolor
Background: Streptomycetes are renowned for their capacity to produce diverse range of pharmaceutically important compounds. Two- thirds of clinically relevant antibiotics are produced by streptomycetes. Beside, these bacteria exhibit a complex and tightly regulated developmental life programme. Single-strand DNA-binding (SSB) proteins have a key role in DNA replication, repair and recombination in all domains of life. SSB act as recruitment platform for genome maintenance proteins and modulates their activities. Many bacteria have additional copies of ssb genes, but their biological role is poorly understood. We found that paralogous Strepomyces coelicolor SsbA is essential while SsbB participate in chromosomal segregation during sporulation. Their affinity for ssDNA was not examined in detail until now. Objectives: To analyze in detail interaction of SsbA and SsbB with single-stranded DNA and to examine localization of these paralogous proteins during S. coelicolor growth. Methods: Genes encoding for His-tagged SsbA and SsbB proteins were cloned into high number copy plasmids. SSBs were expressed in E. coli and purified using affinity and ion-exchange chromatography. EMSA and fluorescence spectroscopy were used to analyze SSBs affinity to ssDNA. Various genetic constructs encoding for a fused GFP or mCherry SSBs were prepared to analyse localization of these proteins in hyphae during bacterial growth. Results: The shortest oligonucleotide bound by SsbA and SsbB was determined under different binding conditions. SsbA:ssDNA was found to be much more dependent on salt concetration compared to SsbB:ssDNA. Localization and occasional co-localization of fluorescently labelled SSB proteins were observed in the vegetative and aerial hyphae of S. coelicolor.
SSB ; Streptomycetes ; localization ; DNA binding ; microscopy
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Podaci o prilogu
1059-1059.
2019.
nije evidentirano
objavljeno
Podaci o matičnoj publikaciji
FEMS2019 : abstract book
Podaci o skupu
8th Congress of European microbiologists (FEMS2019)
poster
07.07.2019-11.07.2019
Glasgow, Ujedinjeno Kraljevstvo