Naslov
DNA binding properties and localization of paralogous SSB proteins from Streptomyces coelicolor

Autori
Tina Paradžik ; Vlatka Godinić Mikulčić ; Anja Kostelac ; Dušica Vujaklija

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

Izvornik
FEMS2019 : abstract book / - , 2019, 1059-1059

Skup
8th Congress of European microbiologists (FEMS2019)

Mjesto i datum
Glasgow, Ujedinjeno Kraljevstvo, 07.07.2019. - 11.07.2019

Vrsta sudjelovanja
Poster

Vrsta recenzije
Međunarodna recenzija

Ključne riječi
SSB ; Streptomycetes ; localization ; DNA binding ; microscopy

Sažetak
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.

Izvorni jezik
Engleski

Znanstvena područja
Biologija

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