Involvement of lysosomal integral membrane protein-2 in the activation of autophagy

Lysosomal integral membrane protein-2 (LIMP-2) is a type III transmembrane protein that is highly glycosylated and mainly localized to the lysosomal membrane. The diverse functions of LIMP-2 are currently being uncovered; however, its participation in macroautophagy, usually described as autophagy,...

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Published in:	Biochemical and biophysical research communications Vol. 533; no. 4; pp. 976 - 982
Main Authors:	Sakane, Hiroshi, Urabe, Junna, Nakahira, Saki, Hino, Katsumi, Miyata, Nao, Akasaki, Kenji
Format:	Journal Article
Language:	English
Published:	United States Elsevier Inc 17-12-2020
Subjects:	Animals Autophagosomes - metabolism Autophagy - physiology CD36 Antigens - antagonists & inhibitors CD36 Antigens - genetics CD36 Antigens - metabolism Chlorocebus aethiops COS Cells Gene Knockdown Techniques HEK293 Cells Humans LC3-II LIMP-2 Lysosomal Membrane Proteins - antagonists & inhibitors Lysosomal Membrane Proteins - genetics Lysosomal Membrane Proteins - metabolism Lysosomes Lysosomes - metabolism Macroautophagy Mice Microtubule-Associated Proteins - genetics Microtubule-Associated Proteins - metabolism NIH 3T3 Cells Rats Recombinant Proteins - genetics Recombinant Proteins - metabolism RNA, Messenger - genetics RNA, Messenger - metabolism Lysosomes LC3-II LIMP-2 Macroautophagy
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Summary:	Lysosomal integral membrane protein-2 (LIMP-2) is a type III transmembrane protein that is highly glycosylated and mainly localized to the lysosomal membrane. The diverse functions of LIMP-2 are currently being uncovered; however, its participation in macroautophagy, usually described as autophagy, has not yet been well-investigated. To determine the possible involvement of LIMP-2 in autophagic activity, we examined the intracellular amount of microtubule-associated protein 1 light chain 3 (LC3)-II, which is well-correlated with autophagosome levels, in exogenous rat LIMP-2-expressing COS7 and HEK293 cells. Transient or stable expression of LIMP-2-myc significantly increased the levels of LC3-II. Conversely, knockdown of LIMP-2 decreased the LC3-II levels in NIH3T3 cells. Furthermore, approaches using lysosomal protease inhibitors and mCherry-GFP-LC3 fluorescence suggested that exogenous expression of LIMP-2 increased the biogenesis of autophagosomes rather than decreased the lysosomal turnover of LC3-II. Considering the results of the biochemical assay and the quantitative fluorescence assay together, it is suggested that LIMP-2 has a possible involvement in autophagic activity, especially autophagosome biogenesis. •Exogenous expression of LIMP-2 increases LC3-II protein level.•Knockdown of LIMP-2 decreases LC3-II protein level.•Approaches using protease inhibitors and mCherry-GFP-LC3 indicate the involvement of LIMP-2 in biogenesis of autophagosomes.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	0006-291X 1090-2104
DOI:	10.1016/j.bbrc.2020.09.114