Internally ratiometric fluorescent sensors for evaluation of intracellular GTP levels and distribution

Bianchi‐Smiraglia, Anna; Rana, Mitra S.; Foley, C E; Paul, L Miller; Lipchick, Brittany C.; Moparthy, Sudha; Moparthy, Kalyana; Fink, Emily E.; Bagati, Archis; Hurley, Edward; Affronti, Hayley C.; Bakin, Andrei V.; Kandel, Eugene; Smiraglia, Dominic J.; Feltri, M. Laura; Sousa, Rui; Nikiforov, Mikhail A.

doi:10.1038/nmeth.4404

Cited by 46 publications

(37 citation statements)

References 33 publications

(45 reference statements)

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“…It would be interesting to investigate Miro-GTP levels in the presence of WT DISC1 or mutDISC1 in live cells. Coupling a GTP/GDP sensor (as described in Bianchi-Smiraglia et al, 2017) to Miro would allow live detection of the bound nucleotide and could even be correlated with the direction of transport-e.g., are anterogradely moving mitochondria likely to be labeled with Miro-GTP? The effect of perturbing DISC1 expression could thus be readily studied.…”

Section: Discussionmentioning

confidence: 99%

DISC1 Regulates Mitochondrial Trafficking in a Miro1-GTP-Dependent Manner

Norkett

Lesept

Kittler

2020

Front. Cell Dev. Biol.

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The disrupted in schizophrenia 1 (DISC1) protein is implicated in major mental illnesses including schizophrenia and bipolar disorder. A key feature of psychiatric disease is aberrant synaptic communication. Correct synaptic transmission is dependent on spatiotemporally regulated energy provision and calcium buffering. This can be achieved by precise distribution of mitochondria throughout the elaborate architecture of the neuron. Central to this process is the calcium sensor and GTPase Miro1, which allows mitochondrial trafficking by molecular motors. While the role of Miro1-calcium binding in mitochondrial transport is well described, far less is known regarding the functions of the two GTPase domains. Here, we investigate the effects of a psychiatric diseaseassociated mutation in DISC1 on mitochondrial trafficking. We show that this DISC1 mutation impairs Miro1's ability to transport mitochondria. We also demonstrate the necessity of the first Miro1 GTPase domain in determining direction of mitochondrial transport and the involvement of DISC1 in this process. Finally, we describe the effects of mutant DISC1 on positioning of mitochondria at synapses.

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Section: Discussionmentioning

confidence: 99%

DISC1 Regulates Mitochondrial Trafficking in a Miro1-GTP-Dependent Manner

Norkett

Lesept

Kittler

2020

Front. Cell Dev. Biol.

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“…For the GTP enrichment model, the visualization of GTP concentration is vital. Recently, a circularly permutated YFP (cpYFP)-based GTP sensor was developed [ 146 ]. GTP imaging using the cpYFP could open another door in the field of NDPK protein and dynamin protein research.…”

Section: Discussionmentioning

confidence: 99%

Molecular Basis of Mitochondrial and Peroxisomal Division Machineries

Imoto

Itoh

Fujiki

2020

IJMS

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Mitochondria and peroxisomes are ubiquitous subcellular organelles that are highly dynamic and possess a high degree of plasticity. These organelles proliferate through division of pre-existing organelles. Studies on yeast, mammalian cells, and unicellular algae have led to a surprising finding that mitochondria and peroxisomes share the components of their division machineries. At the heart of the mitochondrial and peroxisomal division machineries is a GTPase dynamin-like protein, Dnm1/Drp1, which forms a contractile ring around the neck of the dividing organelles. During division, Dnm1/Drp1 functions as a motor protein and constricts the membrane. This mechanochemical work is achieved by utilizing energy from GTP hydrolysis. Over the last two decades, studies have focused on the structure and assembly of Dnm1/Drp1 molecules around the neck. However, the regulation of GTP during the division of mitochondrion and peroxisome is not well understood. Here, we review the current understanding of Dnm1/Drp1-mediated divisions of mitochondria and peroxisomes, exploring the mechanisms of GTP regulation during the Dnm1/Drp1 function, and provide new perspectives on their potential contribution to mitochondrial and peroxisomal biogenesis.

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“…Thus, expected analyte concentration also needs to be considered when selecting the best version of biosensors. In some cases, selecting versions with different kinetic constants or affinity is beneficial as different situations require probes with varying sensitivity [82,87,88,89,97,98,99,100,101,102].…”

Section: General Principles For Developing Cpfp-based Gefismentioning

confidence: 99%

Circularly Permuted Fluorescent Protein-Based Indicators: History, Principles, and Classification

Kostyuk

Demidovich

Kotova

et al. 2019

IJMS

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Genetically encoded biosensors based on fluorescent proteins (FPs) are a reliable tool for studying the various biological processes in living systems. The circular permutation of single FPs led to the development of an extensive class of biosensors that allow the monitoring of many intracellular events. In circularly permuted FPs (cpFPs), the original N- and C-termini are fused using a peptide linker, while new termini are formed near the chromophore. Such a structure imparts greater mobility to the FP than that of the native variant, allowing greater lability of the spectral characteristics. One of the common principles of creating genetically encoded biosensors is based on the integration of a cpFP into a flexible region of a sensory domain or between two interacting domains, which are selected according to certain characteristics. Conformational rearrangements of the sensory domain associated with ligand interaction or changes in the cellular parameter are transferred to the cpFP, changing the chromophore environment. In this review, we highlight the basic principles of such sensors, the history of their creation, and a complete classification of the available biosensors.

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Internally ratiometric fluorescent sensors for evaluation of intracellular GTP levels and distribution

Cited by 46 publications

References 33 publications

DISC1 Regulates Mitochondrial Trafficking in a Miro1-GTP-Dependent Manner

DISC1 Regulates Mitochondrial Trafficking in a Miro1-GTP-Dependent Manner

Molecular Basis of Mitochondrial and Peroxisomal Division Machineries

Circularly Permuted Fluorescent Protein-Based Indicators: History, Principles, and Classification

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