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Differential roles of N- and C-terminal LIR motifs in the catalytic activity and membrane targeting of RavZ and ATG4B proteins
Sang-Won Park 1,2 (Graduate student), Ju-Hui Park3 (Graduate student), Haneul Choi4 (Graduate student), Pureum Jeon4 (Graduate student), Seung-Hwan Lee3 (Graduate student), Won-Dong Shin1,2 (Graduate student), Hun-Joo Kim1 (Graduate student), Jin-A Lee 4 (Professor), Deok-Jin Jang 1,3,* (Professor)
1Department of Vector Entomology, College of Ecology and Environment, Kyungpook National University, 2559, Gyeongsang-daero, Sangju-si, Gyeongsangbuk-do 37224, Republic of Korea,
2Research Institute of Invertebrate Vector, Kyungpook National University, Sangju, Korea,
3Department of Ecological Science, College of Ecology and Environment, Kyungpook National University, 2559, Gyeongsang-daero, Sangju-si, Gyeongsangbuk-do 37224, Republic of Korea,
4Department of Biological Sciences and Biotechnology, College of Life Sciences and Nanotechnology, Hannam University, 1646, Yuseong-daero, Yuseong-gu, Daejeon 34054, Republic of Korea
Abstract
Mammalian ATG8 proteins (mATG8s) are essential for selective autophagy because they recruit various proteins with LC3-interacting region (LIR) motifs to autophagic membranes. The RavZ protein, secreted by Legionella pneumophila, and mammalian ATG4B possess functional LIR motifs that participate in lipidated mATG8 deconjugation on autophagic membranes. RavZ comprises three functional LIR motifs at the N- and C-terminal sides of its catalytic domain (CAD). This study demonstrated that LIR motifs at the N-terminal side of the CAD of RavZ are involved in autophagic membrane targeting and substrate recognition, while LIR motif at the C-terminal side facilitate autophagic membrane targeting. Our results also revealed that the C-terminal LIR motif in human ATG4B is pivotal in delipidating LC3B-phosphatidylethanolamine (PE), but it plays a minor role in pro-LC3B priming in the cytosol. Therefore, introducing a functional LIR motif to the N-terminal of ATG4B does not affect LC3B-PE delipidation. This study clearly described the position-dependent roles of LIR motifs in RavZ and ATG4B in cellular contexts.
Abstract, Accepted Manuscript [Submitted on May 27, 2024, Accepted on September 2, 2024]
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