Naslov
Binding properties and localization of paralogous SSB proteins in hyphae of Streptomyces coelicolor

Autori
Paradžik, Tina ; Godinić Mikulčić, Vlatka ; Kosalec, Anja ; Vujaklija, Dušica

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, ostalo

Izvornik
Book of abstracts of Power of microbes in industry and environment 2019 / - , 2019, 60-60

Skup
Power of Microbes in Industry and Environment 2019

Mjesto i datum
Sveti Martin na Muri, Hrvatska, 15.05.2019. - 18.05.2019

Vrsta sudjelovanja
Poster

Vrsta recenzije
Nije recenziran

Ključne riječi
DNA binding ; SSB ; localization ; EMSA ; mycelia

Sažetak
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.

Izvorni jezik
Engleski

Znanstvena područja
Biologija

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