Binding properties and localization of paralogous SSB proteins in hyphae of Streptomyces coelicolor (CROSBI ID 695329)
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Podaci o odgovornosti
Paradžik, Tina ; Godinić Mikulčić, Vlatka ; Kosalec, Anja ; Vujaklija, Dušica
engleski
Binding properties and localization of paralogous SSB proteins in hyphae of Streptomyces coelicolor
Streptomycetes are complex bacteria which are known as potent producers of various pharmacologically important compounds such as antibiotics, immunosuppressants and anticancer drugs. 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. ssDNA-SSB act as recruitment platform for many genome maintenance proteins. While doing so, it modulates their activities. Although many bacteria have additional copies of ssb genes, their biological role is poorly understood. We previously found that in Strepomyces coelicolor SsbA is essential while SsbB participate in chromosomal segregation during sporulation. Their affinity for binding to ssDNA was not examined in detail. Objectives of this study were (a) to explore SsbA and SsbB interaction with single-stranded DNA and (b) to examine localization of these paralogous proteins during S. coelicolor growth. Genes encoding for SsbA and SsbB protein were His-tagged and cloned on the high number copy plasmids. They were expressed in E.coli and purified using affinity and ion-exchange chromatography. Elecrophoretic mobility shift assay (EMSA) and fluorescence spectroscopy were used to analyse SsbA and SsbB affinity to ssDNA. Various genetic constructs were prepared to analyse localization of these proteins under confocal microscope. The shortest oligonucleotide bound by SsbA and SsbB with a respect to salt concentration was determined. SsbA:ssDNA was found to be much more dependent on salt concetration compared to SsbB:ssDNA. Localisation of fluorescently tagged SSB proteins was observed in the vegetative and aerial hyphae.
DNA binding ; SSB ; localization ; EMSA ; mycelia
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Podaci o prilogu
60-60.
2019.
nije evidentirano
objavljeno
Podaci o matičnoj publikaciji
Book of abstracts of Power of microbes in industry and environment 2019
Podaci o skupu
Power of Microbes in Industry and Environment 2019
poster
15.05.2019-18.05.2019
Sveti Martin na Muri, Hrvatska