Time-Resolved Detection of Light-Induced Dimerization of Monomeric Aureochrome-1 and Change in Affinity for DNA

Akiyama, Yuki; Nakasone, Yusuke; Nakatani, Yôichi; Hisatomi, Osamu; Terazima, Masahide

doi:10.1021/acs.jpcb.6b05760

Cited by 27 publications

(36 citation statements)

References 44 publications

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“…In the dark, PtAUREO1a has been observed as a monomer in equilibrium with a dimer ( Heintz and Schlichting, 2016 ) or as a dimer/higher oligomer ( Banerjee et al., 2016b ). Light induces the dimerization of LOV domains and the association of the monomers ( Herman et al., 2013 ; Goett-Zink et al., 2020 ; Kobayashi et al., 2020 ), which is the rate-limiting step in the process of DNA binding ( Akiyama et al., 2016 ). The diatom P. tricornutum contains four genes encoding different aureochromes ( Schellenberger Costa et al., 2013 ).…”

Section: Introductionmentioning

confidence: 99%

The Aureochrome Photoreceptor PtAUREO1a Is a Highly Effective Blue Light Switch in Diatoms

Mann¹,

Serif²,

Wrobel³

et al. 2020

iScience

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Summary Aureochromes represent a unique type of blue light photoreceptors that possess a blue light sensing flavin-binding LOV-domain and a DNA-binding bZIP domain, thus being light-driven transcription factors. The diatom Phaeodactylum tricornutum , a member of the essential marine primary producers, possesses four aureochromes (PtAUREO1a, 1b, 1c, 2). Here we show a dramatic change in the global gene expression pattern of P. tricornutum wild-type cells after a shift from red to blue light. About 75% of the genes show significantly changed transcript levels already after 10 and 60 min of blue light exposure, which includes genes of major transcription factors as well as other photoreceptors. Very surprisingly, this light-induced regulation of gene expression is almost completely inhibited in independent PtAureo1a knockout lines. Such a massive and fast transcriptional change depending on one single photoreceptor is so far unprecedented. We conclude that PtAUREO1a plays a key role in diatoms upon blue light exposure.

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Section: Introductionmentioning

confidence: 99%

The Aureochrome Photoreceptor PtAUREO1a Is a Highly Effective Blue Light Switch in Diatoms

Mann¹,

Serif²,

Wrobel³

et al. 2020

iScience

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“…Dynamic light scattering and size-exclusion chromatography (SEC) analyses showed that recombinant Vf AUREO1 is monomeric in reducing conditions in the dark state [ 23 ]. BL induces non-covalent dimerization, which increases its affinity for the target DNA sequence [ 23 , 24 ]. These results suggest that dimerization has an important role in DNA binding by Vf AUREO1.…”

mentioning

confidence: 99%

Quantitative analyses of the equilibria among DNA complexes of a blue-light-regulated bZIP module, Photozipper

Nakatani

Hisatomi

2018

BIOPHYSICS

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Aureochrome1 is a blue-light-receptor protein identified in a stramenopile alga, Vaucheria frigida. Photozipper (PZ) is an N-terminally truncated, monomeric, V. frigida aureochrome1 fragment containing a basic leucine zipper (bZIP) domain and a light–oxygen–voltage (LOV)-sensing domain. PZ dimerizes upon photoexcitation and consequently increases its affinity for the target sequence. In the present study, to understand the equilibria among DNA complexes of PZ, DNA binding by PZ and mutational variants was quantitatively investigated by electrophoretic-mobility-shift assay and fluorescence-correlation spectroscopy in the dark and light states. DNA binding by PZ was sequence-specific and light-dependent. The half-maximal effective concentration of PZ for binding to the target DNA sequence was ~40 nM in the light, which was >10-fold less than the value in the dark. By contrast, the dimeric PZ-S2C variant (with intermolecular disulfide bonds) had higher affinity for the target sequence, with dissociation constants of ~4 nM, irrespective of the light conditions. Substitutions of Glu159 and Lys164 in the leucine zipper region decreased the affinity of PZ for the target sequence, especially in the light, suggesting that these residues form inter-helical salt bridges between leucine zipper regions, stabilizing the dimer–DNA complex. Our quantitative analyses of the equilibria in PZ–DNA-complex formation suggest that the blue-light-induced dimerization of LOV domains and coiled-coil formation by leucine zipper regions are the primary determinants of the affinity of PZ for the target sequence.

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“…However, the intermolecular interaction dynamics and the recognition mechanism of the target sequence had remained unclear. Hence, the photoreaction dynamics of EL222 and Aureo1 in the absence and presence of DNA were studied by the TG method (Akiyama et al, 2016;Takakado et al, 2017Takakado et al, , 2018.…”

Section: Studies On Photoreactive Proteinsmentioning

confidence: 99%

A Time-Resolved Diffusion Technique for Detection of the Conformational Changes and Molecular Assembly/Disassembly Processes of Biomolecules

Nakasone

Terazima

2021

Front. Genet.

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Biological liquid–liquid phase separation (LLPS) is driven by dynamic and multivalent interactions, which involves conformational changes and intermolecular assembly/disassembly processes of various biomolecules. To understand the molecular mechanisms of LLPS, kinetic measurements of the intra- and intermolecular reactions are essential. In this review, a time-resolved diffusion technique which has a potential to detect molecular events associated with LLPS is presented. This technique can detect changes in protein conformation and intermolecular interaction (oligomer formation, protein-DNA interaction, and protein-lipid interaction) in time domain, which are difficult to obtain by other methods. After the principle and methods for signal analyses are described in detail, studies on photoreactive molecules (intermolecular interaction between light sensor proteins and its target DNA) and a non-photoreactive molecule (binding and folding reaction of α-synuclein upon mixing with SDS micelle) are presented as typical examples of applications of this unique technique.

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Time-Resolved Detection of Light-Induced Dimerization of Monomeric Aureochrome-1 and Change in Affinity for DNA

Cited by 27 publications

References 44 publications

The Aureochrome Photoreceptor PtAUREO1a Is a Highly Effective Blue Light Switch in Diatoms

The Aureochrome Photoreceptor PtAUREO1a Is a Highly Effective Blue Light Switch in Diatoms

Quantitative analyses of the equilibria among DNA complexes of a blue-light-regulated bZIP module, Photozipper

A Time-Resolved Diffusion Technique for Detection of the Conformational Changes and Molecular Assembly/Disassembly Processes of Biomolecules

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