Photo-dependent membrane-less organelles formed from plant phyB and PIF6 proteins in mammalian cells

Fonin, Alexander V.; Antifeeva, Iuliia A.; Shpironok, Olesya G.; Stepanenko, Olesya V.; Silonov, Sergey A.; Antifeev, Ivan; Romanovich, Anna E.; Kuznetsova, Irina М.; Kim, Jeong-Il; Uversky, Vladimir N.; Turoverov, Konstantin К.

doi:10.1016/j.ijbiomac.2021.02.075

Cited by 9 publications

(5 citation statements)

References 37 publications

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“…Although PIF4 can also accumulate in the light 36,45,54 , PIF4 is ubiquitylated by the cullin3-based E3 ligase CRL3 BOP1/2 using BLADE ON PETIOLE (BOP1/2) as the substrate recognition components 76 . When coexpressed with PHYB, FP-tagged PIF4, as well as PIF3 and PIF6/PIL2, localized with PHYB in biomolecular condensates in mammalian cells or protoplasts in a PHYB-dependent manner, supporting the idea that PHYB may be able to recruit PIF4 to PBs [37][38][39] . However, the PB localization of PIF4 in Arabidopsis has not been carefully studied, and the potential role of PBs in stabilizing PIF4 in the light remains to be experimentally verified.…”

Section: Discussionmentioning

confidence: 59%

“…A D1040V mutation in this module, which disrupts PHYB dimerization, abolishes PHYB's ability to form PBs 36 . Fourth, PHYB can undergo light-dependent LLPS into biomolecular condensates in mammalian cells [37][38][39] . The intramolecular attributes of PHYB required for PB formation in vivo, e.g., the C-terminal module and N-terminal extension, are also essential for PHYB LLPS in this heterologous system 37 , supporting the idea that PB formation in vivo may be propelled by the LLPS of active PHYB alone.…”

Section: Introductionmentioning

confidence: 99%

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Photobody formation spatially segregates two opposing phytochrome B signaling actions to titrate plant environmental responses

Kim,

Fan,

et al. 2023

Preprint

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Photoactivation of the plant photoreceptor and thermosensor phytochrome B (PHYB) triggers its condensation into subnuclear photobodies (PBs). However, the function of PBs remains frustratingly elusive. Here, we show that PHYB condensation enables the co-occurrence and competition of two antagonistic phase-separated signaling actions. We found that PHYB recruits PHYTOCHROME-INTERACTING FACTOR5 (PIF5) to PBs and, surprisingly, that PHYB exerts opposing roles in degrading and stabilizing PIF5. Perturbing PB size by overproducing PHYB provoked a biphasic PIF5 response: while a moderate increase in PHYB enhanced PIF5 degradation, further elevating the PHYB level stabilized PIF5 by retaining more of it in enlarged PBs. Our results support a model in which PHYB condensation stabilizes PIF5 in PBs to counteract PIF5 degradation in the surrounding nucleoplasm, thereby enabling an environmentally sensitive counterbalancing mechanism to titrate nucleoplasmic PIF5 and its transcriptional output. This PB-enabled signaling mechanism provides a framework for regulating a plethora of PHYB-interacting signaling molecules in diverse plant environmental responses. We propose that this function of PBs represents a general function of biomolecular condensates to allow distinct variations of a cellular process or signaling pathway to coexist and interact to generate dynamically adjustable integrated outputs within a single subcellular space.

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Section: Discussionmentioning

confidence: 59%

Section: Introductionmentioning

confidence: 99%

Photobody formation spatially segregates two opposing phytochrome B signaling actions to titrate plant environmental responses

Kim,

Fan,

et al. 2023

Preprint

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“…Naturally existing photoreceptors respond to different wavelengths of light, most of which were blue light and red/far-red light receptors. Reported red/far-red absorbing photoreceptors, including Cph1 (85 kDa), llaC (85 kDa), PhyB (130 kDa), and BphP1 (80 kDa), belong to the phytochromes family. − Red/far-red photoreceptors were usually fused with transcriptional regulators to photocontrol the transcription of certain genes. However, the molecular weight of red/far-red photosensors is too large to insert into a protein because when a larger photoreceptor is inserted, the structure of the IDH is further disturbed.…”

Section: Discussionmentioning

confidence: 99%

Photocontrol of Itaconic Acid Synthesis in Escherichia coli

Zhao

Wei

et al. 2022

ACS Synth. Biol.

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Metabolic engineering aims to control cellular metabolic flow and maximize the production of a product of interest. Photocontrol of the activities of proteins is an effective method for accurately regulating metabolic pathways. In this study, we inserted the photosensor light-oxygen-voltage-sensing domain 2 of Avena sativa (AsLOV2) into selected sites of isocitrate dehydrogenase (IDH), the key enzyme in the competitive pathway of itaconic acid (ITA) synthesis, to construct photoswitchable IDH-AsLOV2 (ILOVs). These engineered light-sensitive proteins were used to regulate the metabolic flux of the tricarboxylic acid (TCA) cycle in Escherichia coli to improve ITA production. The engineered fusion proteins ILOV2, ILOV3, ILOV6, and ILOV7 exhibited effective reversibility under the oscillation of darkness and blue light illumination in vitro. The efficacies of the intracellular photoswitches were evaluated, and an optimal photocontrol strategy was established in vivo. The ITA titer was significantly enhanced to 3.30 g/L for strain ITAΔ43, which displayed superior photoswitchable potency for ITA production compared with the strains that completely deleted the icd gene. The photocontrol strategy developed here can be extended for process optimization and titer improvement of other high-value bioengineering chemicals.

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“…The first systems were based on the plant phytochrome B and its binding partner Phytochrome-Interacting Factor (PIF) ( Shimizu-Sato et al, 2002 ). These systems are still routinely utilized despite the hindrance of their larger size and complicated application due to the non-mammalian chromophore ( Raghavan et al, 2020 ; Fonin et al, 2021 ). However, successful efforts have been made to decrease the size of PIF and increase its binding affinity to phytochrome ( Golonka et al, 2019 ; Yousefi et al, 2019 ).…”

Section: Red Light-sensing Optogenetic Actuatorsmentioning

confidence: 99%

Red Light Optogenetics in Neuroscience

Lehtinen

Nokia

Takala

2022

Front. Cell. Neurosci.

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Optogenetics, a field concentrating on controlling cellular functions by means of light-activated proteins, has shown tremendous potential in neuroscience. It possesses superior spatiotemporal resolution compared to the surgical, electrical, and pharmacological methods traditionally used in studying brain function. A multitude of optogenetic tools for neuroscience have been created that, for example, enable the control of action potential generation via light-activated ion channels. Other optogenetic proteins have been used in the brain, for example, to control long-term potentiation or to ablate specific subtypes of neurons. In in vivo applications, however, the majority of optogenetic tools are operated with blue, green, or yellow light, which all have limited penetration in biological tissues compared to red light and especially infrared light. This difference is significant, especially considering the size of the rodent brain, a major research model in neuroscience. Our review will focus on the utilization of red light-operated optogenetic tools in neuroscience. We first outline the advantages of red light for in vivo studies. Then we provide a brief overview of the red light-activated optogenetic proteins and systems with a focus on new developments in the field. Finally, we will highlight different tools and applications, which further facilitate the use of red light optogenetics in neuroscience.

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Photo-dependent membrane-less organelles formed from plant phyB and PIF6 proteins in mammalian cells

Cited by 9 publications

References 37 publications

Photobody formation spatially segregates two opposing phytochrome B signaling actions to titrate plant environmental responses

Photobody formation spatially segregates two opposing phytochrome B signaling actions to titrate plant environmental responses

Photocontrol of Itaconic Acid Synthesis in Escherichia coli

Red Light Optogenetics in Neuroscience

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