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Functional and structural insights into one-component transcriptional regulators involved in methylenomycin production in Streptomyces coelicolor A3(2)
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Ajaykumar, Rohini (2021) Functional and structural insights into one-component transcriptional regulators involved in methylenomycin production in Streptomyces coelicolor A3(2). PhD thesis, University of Warwick.
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Official URL: http://webcat.warwick.ac.uk/record=b3821713~S15
Abstract
In one-component regulatory systems a transcriptional regulator (TR) protein
typically comprises both ligand and DNA binding domains to prevent or promote
the expression of target gene(s). One component TRs are present in the
methylenomycin (an antibiotic) gene cluster in Streptomyces coelicolor A3(2). These
are involved in controlling methylenomycin biosynthesis by the TR MmyB and
controlling methylenomycin resistance by the TR MmyJ. The characterisation of
these two TRs was the focus of this thesis.
Bioinformatic analysis of MmyJ indicate it to be an ArsR family (a metal sensing
family of TRs) protein with uncharacterised one-component MmyJ like systems
suggested in other actinobacteria. DNA-MmyJ binding assays revealed MmyJ binds
to two 17 base pair pseudo-palindromic regions beside one another within the
intergenic region between mmr (MFS type efflux pump and target gene of MmyJ)
and mmyJ. Bacterial hybrid assays showed MmyJ forming homodimers and
mutation of Leu25 highlighted it as a key residue for dimer formation. X-ray
diffraction of MmyJ meant its structure was solved to 1.68 Å. Structural analysis of
MmyJ corroborates it to be an ArsR family protein meaning MmyJ is the first
antibiotic sensing ArsR family protein structure to be solved.
X-ray diffraction was also used for crystals of MmyB however these did not diffract
sufficiently for structure elucidation. Using liquid chromatography mass
spectrometry, the regulation upon mmyb was able to be evaded in order for
methylenomycin biosynthesis, highlighting the significance of MmyB. This also
indicates a potential method for discovery of novel secondary metabolites through
over expression of mmyb-like genes.
Item Type: | Thesis (PhD) | ||||
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Subjects: | Q Science > QR Microbiology | ||||
Library of Congress Subject Headings (LCSH): | Streptomyces coelicolor, Streptomyces -- Genetics, Genetic transcription -- Regulation, Drug resistance in microorganisms, Natural products -- Research | ||||
Official Date: | December 2021 | ||||
Dates: |
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Institution: | University of Warwick | ||||
Theses Department: | School of Life Sciences | ||||
Thesis Type: | PhD | ||||
Publication Status: | Unpublished | ||||
Supervisor(s)/Advisor: | Corre, Christophe, Crow, Allister | ||||
Sponsors: | Biotechnology and Biological Sciences Research Council (Great Britain) | ||||
Extent: | xix, 274 leaves : illustrations (colour), charts (colour) | ||||
Language: | eng |
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