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Monitoring LC3-or GABARAP-positive autophagic membranes using modified RavZ-based probes

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Title
Monitoring LC3-or GABARAP-positive autophagic membranes using modified RavZ-based probes
Author(s)
Park, SW; Jeon, P; Jun, YW; Park, JH; Lee, SH; Sangkyu Lee; Lee, JA; Jang, DJ
Publication Date
2019-11
Journal
SCIENTIFIC REPORTS, v.9, no., pp.16593 -
Publisher
NATURE PUBLISHING GROUP
Abstract
Xenophagy is a selective lysosomal degradation pathway for invading pathogens in host cells. However, invading bacteria also develop survival mechanisms to inhibit host autophagy. RavZ is a protein secreted by Legionella that irreversibly delipidates mammalian autophagy-related protein 8 (mATG8) on autophagic membranes in host cells via efficient autophagic membrane targeting. In this study, we leveraged the autophagic membrane-targeting mechanism of RavZ and generated a new autophagosome probe by replacing the catalytic domain of RavZ with GFP. This probe is efficiently localized to mATG8-positive autophagic membranes via a synergistic combination between mATG8 protein-binding mediated by the LC3-interacting region (LIR) motifs and phosphoinositide-3-phosphate (PI3P) binding mediated by the membrane-targeting (MT) domain. Furthermore, the membrane association activity of this new probe with an MT domain was more efficient than that of probes with a hydrophobic domain that were previously used in LIR-based autophagosome sensors. Finally, by substituting the LIR motifs of RavZ with selective LIR motifs from Fycol or ULK2, we developed new probes for detecting LC3A/B- or GABARAP subfamily-positive autophagic membranes, respectively. We propose that these new RavZ-based sensors will be useful for monitoring and studying the function of mATG8-positive autophagic membranes in different cellular contexts for autophagy research. © The Author(s) 2019
URI
https://pr.ibs.re.kr/handle/8788114/6776
ISSN
2045-2322
Appears in Collections:
Center for Cognition and Sociality(인지 및 사회성 연구단) > Social Neuroscience Group(사회성 뇌과학 그룹) > Journal Papers (저널논문)
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[2019]Sci.Rep.-Monitoring LC3- or GABARAP positive autophagic membranes_RavZ probes.pdfDownload

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