The Photoconvertible Fluorescent Protein Dendra2 Tag as a Tool to Investigate Intracellular Protein Dynamics

Lešková, Alexandra; Kusá, Zuzana; Labajová, Mária; Krausko, Miroslav; Jásik, Ján

doi:10.1007/978-1-4939-9469-4_13

Cited by 6 publications

(5 citation statements)

References 38 publications

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“…(2013) and Lešková et al. (2019), all root apices of these seedlings were then photoconverted with blue light at 405 nm for 1 min using 30% intensity of CSLM. Subsequently, the converted roots were treated with solutions containing 0 or 50 μ m H 3 BO 3 and 0 or 50 m m CHX at pH 5.8 for different time periods ranging from 0 to 4 h. Real‐time in vivo analysis of Dendra2 signals in the same positions was performed using CSLM.…”

Section: Methodsmentioning

confidence: 99%

PIN2/3/4 auxin carriers mediate root growth inhibition under conditions of boron deprivation in Arabidopsis

et al. 2023

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SUMMARYThe mechanistic basis by which boron (B) deprivation inhibits root growth via the mediation of root apical auxin transport and distribution remains elusive. This study showed that B deprivation repressed root growth of wild‐type Arabidopsis seedlings, which was related to higher auxin accumulation (observed with DII‐VENUS and DR5‐GFP lines) in B‐deprived roots. Boron deprivation elevated the auxin content in the root apex, coinciding with upregulation of the expression levels of auxin biosynthesis‐related genes (TAA1, YUC3, YUC9, and NIT1) in shoots, but not in root apices. Phenotyping experiments using auxin transport‐related mutants revealed that the PIN2/3/4 carriers are involved in root growth inhibition caused by B deprivation. B deprivation not only upregulated the transcriptional levels of PIN2/3/4, but also restrained the endocytosis of PIN2/3/4 carriers (observed with PIN‐Dendra2 lines), resulting in elevated protein levels of PIN2/3/4 in the plasma membrane. Overall, these results suggest that B deprivation not only enhances auxin biosynthesis in shoots by elevating the expression levels of auxin biosynthesis‐related genes but also promotes the polar auxin transport from shoots to roots by upregulating the gene expression levels of PIN2/3/4, as well as restraining the endocytosis of PIN2/3/4 carriers, ultimately resulting in auxin accumulation in root apices and root growth inhibition.

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Section: Methodsmentioning

confidence: 99%

PIN2/3/4 auxin carriers mediate root growth inhibition under conditions of boron deprivation in Arabidopsis

et al. 2023

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“…It is likely that this technology can also trace a putative simultaneous release of DNA and DNA-binding proteins from the organelles. State-of-the-art microscopic techniques with augmented resolution have been tested for life-cell imaging in plants, which entails specific challenges such as high light scattering [165,166]. Furthermore, photoconvertible fluorescent proteins such as Dendra2 are emerging as useful tools in tracing a fraction of the whole protein cellular population on its way from an organelle to the nucleus [167,168].…”

Section: Methods Suited For Analyses Of Organelle-nucleus Translocation (A) Strategies Used For Analysis Of the Dual Localization Of Selementioning

confidence: 99%

Genome communication in plants mediated by organelle–nucleus-located proteins

Krupinska

Blanco

Oetke

et al. 2020

Phil. Trans. R. Soc. B

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An increasing number of eukaryotic proteins have been shown to have a dual localization in the DNA-containing organelles, mitochondria and plastids, and/or the nucleus. Regulation of dual targeting and relocation of proteins from organelles to the nucleus offer the most direct means for communication between organelles as well as organelles and nucleus. Most of the mitochondrial proteins of animals have functions in DNA repair and gene expression by modelling of nucleoid architecture and/or chromatin. In plants, such proteins can affect replication and early development. Most plastid proteins with a confirmed or predicted second location in the nucleus are associated with the prokaryotic core RNA polymerase and are required for chloroplast development and light responses. Few plastid–nucleus-located proteins are involved in pathogen defence and cell cycle control. For three proteins, it has been clearly shown that they are first targeted to the organelle and then relocated to the nucleus, i.e. the nucleoid-associated proteins HEMERA and Whirly1 and the stroma-located defence protein NRIP1. Relocation to the nucleus can be experimentally demonstrated by plastid transformation leading to the synthesis of proteins with a tag that enables their detection in the nucleus or by fusions with fluoroproteins in different experimental set-ups. This article is part of the theme issue ‘Retrograde signalling from endosymbiotic organelles’.

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“…Fusion with the Dendra2 allows for non-invasive tracking of endogenous TDP-43 protein levels in live cells via optical pulse labeling (OPL), a technique we previously used to track the turnover of overexpressed TDP-43 in primary neurons 74 . Briefly and as previously described 74 , in OPL we exploit the property of Dendra2 to irreversibly switch emission maxima from green to red wavelengths upon exposure to short-wavelength light 84,85 (termed "photoconversion," Fig. 5B).…”

Section: Mtmr5 Expression Is Selectively Enhanced In Neuronsmentioning

confidence: 99%

Myotubularin-related phosphatase 5 is a critical determinant of autophagy in neurons

et al. 2022

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The Photoconvertible Fluorescent Protein Dendra2 Tag as a Tool to Investigate Intracellular Protein Dynamics

Cited by 6 publications

References 38 publications

PIN2/3/4 auxin carriers mediate root growth inhibition under conditions of boron deprivation in Arabidopsis

PIN2/3/4 auxin carriers mediate root growth inhibition under conditions of boron deprivation in Arabidopsis

Genome communication in plants mediated by organelle–nucleus-located proteins

Myotubularin-related phosphatase 5 is a critical determinant of autophagy in neurons

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The Photoconvertible Fluorescent Protein Dendra2 Tag as a Tool to Investigate Intracellular Protein Dynamics

Cited by 6 publications

References 38 publications

PIN2/3/4 auxin carriers mediate root growth inhibition under conditions of boron deprivation in Arabidopsis

PIN2/3/4 auxin carriers mediate root growth inhibition under conditions of boron deprivation in Arabidopsis

Genome communication in plants mediated by organelle–n­ucleus-located proteins

Myotubularin-related phosphatase 5 is a critical determinant of autophagy in neurons

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Genome communication in plants mediated by organelle–nucleus-located proteins