Please use this identifier to cite or link to this item: http://hdl.handle.net/2445/127218
Title: Plasticity in oligomerization, operator architecture, and DNA binding in the mode of action of a bacterial B12-based photoreceptor.
Author: Fernández-Zapata, Jesús
Pérez-Castaño, Ricardo
Aranda, Juan
Colizzi, Francesco
Polanco, María Carmen
Orozco López, Modesto
Padmanabhan, S.
Elías-Arnanz, Montserrat
Keywords: Fotoreceptors
Plasticitat
Oligòmers
ADN
Photoreceptors
Plasticity
Oligomers
DNA
Issue Date: 27-Sep-2018
Publisher: American Society for Biochemistry and Molecular Biology
Abstract: Newly discovered bacterial photoreceptors called CarH sense light by using 5′-deoxyadenosylcobalamin (AdoCbl). They repress their own expression and that of genes for carotenoid synthesis by binding in the dark to operator DNA as AdoCbl-bound tetramers, whose light-induced disassembly relieves repression. High-resolution structures of Thermus thermophilus CarHTt have provided snapshots of the dark and light states and have revealed a unique DNA-binding mode whereby only three of four DNA-binding domains contact an operator comprising three tandem direct repeats. To gain further insights into CarH photoreceptors and employing biochemical, spectroscopic, mutational, and computational analyses, here we investigated CarHBm from Bacillus megaterium. We found that apoCarHBm, unlike monomeric apoCarHTt, is an oligomeric molten globule that forms DNA-binding tetramers in the dark only upon AdoCbl binding, which requires a conserved W-X9-EH motif. Light relieved DNA binding by disrupting CarHBm tetramers to dimers, rather than to monomers as with CarHTt. CarHBm operators resembled that of CarHTt, but were larger by one repeat and overlapped with the −35 or −10 promoter elements. This design persisted in a six-repeat, multipartite operator we discovered upstream of a gene encoding an Spx global redox-response regulator whose photoregulated expression links photooxidative and general redox responses in B. megaterium. Interestingly, CarHBm recognized the smaller CarHTt operator, revealing an adaptability possibly related to the linker bridging the DNA- and AdoCbl-binding domains. Our findings highlight a remarkable plasticity in the mode of action of B12-based CarH photoreceptors, important for their biological functions and development as optogenetic tools.
Note: Reproducció del document publicat a: https://doi.org/10.1074/jbc.RA118.004838
It is part of: Journal of Biological Chemistry, 2018, vol. 293, num. 46, p. 17888-17905
Related resource: https://doi.org/10.1074/jbc.RA118.004838
URI: http://hdl.handle.net/2445/127218
ISSN: 0021-9258
Appears in Collections:Articles publicats en revistes (Bioquímica i Biomedicina Molecular)
Articles publicats en revistes (Institut de Recerca Biomèdica (IRB Barcelona))

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