Small GTPase signaling hubs at the surface of cellular membranes in physiology and disease

Cherfils, Jacqueline

doi:10.1002/1873-3468.14607

Cited by 2 publications

References 21 publications

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Small GTPase ActIvitY ANalyzing (SAIYAN) system: A method to detect GTPase activation in living cells

Maeda,

Arakawa,

Komatsu

et al. 2024

Journal of Cell Biology

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Small GTPases are essential in various cellular signaling pathways, and detecting their activation within living cells is crucial for understanding cellular processes. The current methods for detecting GTPase activation using fluorescent proteins rely on the interaction between the GTPase and its effector. Consequently, these methods are not applicable to factors, such as Sar1, where the effector also functions as a GTPase-activating protein. Here, we present a novel method, the Small GTPase ActIvitY ANalyzing (SAIYAN) system, for detecting the activation of endogenous small GTPases via fluorescent signals utilizing a split mNeonGreen system. We demonstrated Sar1 activation at the endoplasmic reticulum (ER) exit site and successfully detected its activation state in various cellular conditions. Utilizing the SAIYAN system in collagen-secreting cells, we discovered activated Sar1 localized both at the ER exit sites and ER–Golgi intermediate compartment (ERGIC) regions. Additionally, impaired collagen secretion confined the activated Sar1 at the ER exit sites, implying the importance of Sar1 activation through the ERGIC in collagen secretion.

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Small GTPase ActIvitY ANalyzing (SAIYAN) system: A method to detect GTPase activation in living cells

Maeda,

Arakawa,

Komatsu

et al. 2024

Journal of Cell Biology

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A Novel Method to Visualize Active Small GTPases Unveils Distinct Sites of Sar1 Activation During Collagen Secretion

Maeda,

Arakawa,

Komatsu

et al. 2024

Preprint

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Small GTPases are essential in various cellular signaling pathways, and detecting their activation within living cells is crucial for understanding cellular processes. Fluorescence resonance energy transfer is widely used to study the interaction between activated small GTPases and their effectors, but it is limited to those with well-defined effectors, excluding Sar1. Here, we present a novel method, SAIYAN (Small GTPase ActIvitY ANalyzing), for detecting the activation of endogenous small GTPases via fluorescent signals utilizing a split mNeonGreen system. We demonstrated Sar1 activation at the endoplasmic reticulum (ER) exit site and successfully detected its activation state in various cellular conditions. Utilizing the SAIYAN system in collagen-secreting cells, we discovered activated Sar1 localized both at ER exit sites and ER-Golgi intermediate compartment (ERGIC) regions. Additionally, impaired collagen secretion led to the confinement of activated Sar1 at the ER exit sites, underscoring the significance of Sar1 activation through the ERGIC in collagen secretion.

show abstract

Small GTPase signaling hubs at the surface of cellular membranes in physiology and disease

Cited by 2 publications

References 21 publications

Small GTPase ActIvitY ANalyzing (SAIYAN) system: A method to detect GTPase activation in living cells

Small GTPase ActIvitY ANalyzing (SAIYAN) system: A method to detect GTPase activation in living cells

A Novel Method to Visualize Active Small GTPases Unveils Distinct Sites of Sar1 Activation During Collagen Secretion

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