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izvor podataka: crosbi

Binding properties and localization of paralogous SSB proteins in hyphae of Streptomyces coelicolor (CROSBI ID 695329)

Prilog sa skupa u zborniku | sažetak izlaganja sa skupa

Paradžik, Tina ; Godinić Mikulčić, Vlatka ; Kosalec, Anja ; Vujaklija, Dušica Binding properties and localization of paralogous SSB proteins in hyphae of Streptomyces coelicolor // Book of abstracts of Power of microbes in industry and environment 2019. 2019. str. 60-60

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

Povezanost rada

Biologija