A kinase translocation reporter reveals real-time dynamics of ERK activity in Drosophila

Extracellular signal-regulated kinase (ERK) lies downstream of a core signalling cascade that controls all aspects of development and adult homeostasis. Recent developments have led to new tools to image and manipulate the pathway. However, visualising ERK activity in vivo with high temporal resolut...
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Autor*in:	Alice C. Yuen [verfasserIn] Anadika R. Prasad [verfasserIn] Vilaiwan M. Fernandes [verfasserIn] Marc Amoyel [verfasserIn]

Format:	E-Artikel
Sprache:	Englisch

Erschienen:	2022

Schlagwörter:	drosophila erk signalling kinase translocation reporter live imaging signalling dynamics

Übergeordnetes Werk:	In: Biology Open - The Company of Biologists, 2012, 11(2022), 5
Übergeordnetes Werk:	volume:11 ; year:2022 ; number:5

Links:	Link aufrufen Link aufrufen Link aufrufen Journal toc

DOI / URN:	10.1242/bio.059364

Katalog-ID:	DOAJ043942415

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callnumber-first	Q - Science
author	Alice C. Yuen
spellingShingle	Alice C. Yuen misc QH301-705.5 misc drosophila misc erk signalling misc kinase translocation reporter misc live imaging misc signalling dynamics misc Science misc Q misc Biology (General) A kinase translocation reporter reveals real-time dynamics of ERK activity in Drosophila
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topic_title	QH301-705.5 A kinase translocation reporter reveals real-time dynamics of ERK activity in Drosophila drosophila erk signalling kinase translocation reporter live imaging signalling dynamics
topic	misc QH301-705.5 misc drosophila misc erk signalling misc kinase translocation reporter misc live imaging misc signalling dynamics misc Science misc Q misc Biology (General)
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title	A kinase translocation reporter reveals real-time dynamics of ERK activity in Drosophila
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title_full	A kinase translocation reporter reveals real-time dynamics of ERK activity in Drosophila
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author_browse	Alice C. Yuen Anadika R. Prasad Vilaiwan M. Fernandes Marc Amoyel
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title_sort	kinase translocation reporter reveals real-time dynamics of erk activity in drosophila
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title_auth	A kinase translocation reporter reveals real-time dynamics of ERK activity in Drosophila
abstract	Extracellular signal-regulated kinase (ERK) lies downstream of a core signalling cascade that controls all aspects of development and adult homeostasis. Recent developments have led to new tools to image and manipulate the pathway. However, visualising ERK activity in vivo with high temporal resolution remains a challenge in Drosophila. We adapted a kinase translocation reporter (KTR) for use in Drosophila, which shuttles out of the nucleus when phosphorylated by ERK. We show that ERK-KTR faithfully reports endogenous ERK signalling activity in developing and adult tissues, and that it responds to genetic perturbations upstream of ERK. Using ERK-KTR in time-lapse imaging, we made two novel observations: firstly, sustained hyperactivation of ERK by expression of dominant-active epidermal growth factor receptor raised the overall level but did not alter the kinetics of ERK activity; secondly, the direction of migration of retinal basal glia correlated with their ERK activity levels, suggesting an explanation for the heterogeneity in ERK activity observed in fixed tissue. Our results show that KTR technology can be applied in Drosophila to monitor ERK activity in real-time and suggest that this modular tool can be further adapted to study other kinases.
abstractGer	Extracellular signal-regulated kinase (ERK) lies downstream of a core signalling cascade that controls all aspects of development and adult homeostasis. Recent developments have led to new tools to image and manipulate the pathway. However, visualising ERK activity in vivo with high temporal resolution remains a challenge in Drosophila. We adapted a kinase translocation reporter (KTR) for use in Drosophila, which shuttles out of the nucleus when phosphorylated by ERK. We show that ERK-KTR faithfully reports endogenous ERK signalling activity in developing and adult tissues, and that it responds to genetic perturbations upstream of ERK. Using ERK-KTR in time-lapse imaging, we made two novel observations: firstly, sustained hyperactivation of ERK by expression of dominant-active epidermal growth factor receptor raised the overall level but did not alter the kinetics of ERK activity; secondly, the direction of migration of retinal basal glia correlated with their ERK activity levels, suggesting an explanation for the heterogeneity in ERK activity observed in fixed tissue. Our results show that KTR technology can be applied in Drosophila to monitor ERK activity in real-time and suggest that this modular tool can be further adapted to study other kinases.
abstract_unstemmed	Extracellular signal-regulated kinase (ERK) lies downstream of a core signalling cascade that controls all aspects of development and adult homeostasis. Recent developments have led to new tools to image and manipulate the pathway. However, visualising ERK activity in vivo with high temporal resolution remains a challenge in Drosophila. We adapted a kinase translocation reporter (KTR) for use in Drosophila, which shuttles out of the nucleus when phosphorylated by ERK. We show that ERK-KTR faithfully reports endogenous ERK signalling activity in developing and adult tissues, and that it responds to genetic perturbations upstream of ERK. Using ERK-KTR in time-lapse imaging, we made two novel observations: firstly, sustained hyperactivation of ERK by expression of dominant-active epidermal growth factor receptor raised the overall level but did not alter the kinetics of ERK activity; secondly, the direction of migration of retinal basal glia correlated with their ERK activity levels, suggesting an explanation for the heterogeneity in ERK activity observed in fixed tissue. Our results show that KTR technology can be applied in Drosophila to monitor ERK activity in real-time and suggest that this modular tool can be further adapted to study other kinases.
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title_short	A kinase translocation reporter reveals real-time dynamics of ERK activity in Drosophila
url	https://doi.org/10.1242/bio.059364 https://doaj.org/article/b01851db000e420883205a8fc9dba3ca http://bio.biologists.org/content/11/5/bio059364 https://doaj.org/toc/2046-6390
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