LC3, an autophagosome marker, can be incorporated into protein aggregates independent of autophagy: caution in the interpretation of LC3 localization.
Autophagy is an intracellular bulk degradation system, through which a portion of the cytoplasm is delivered to lysosomes to be degraded. Microtuble-associated protein light chain 3 (LC3), a mammalian homolog of yeast Atg8, has been used as a specific marker to monitor autophagy. Upon induction of autophagy, LC3 is conjugated to phosphatidylethanolamine and targeted to autophagic membranes. Therefore, changes in LC3 localization have been used to measure autophagy. However, this method has some limitations. In this report, we show that LC3 protein tends to aggregate in an autophagy-independent manner when it is transiently overexpressed by transfection. In addition, LC3 is easily incorporated into intracellular protein aggregates, such as inclusion bodies induced by polyQ expression or formed in autophagy-deficient hepatocytes, neurons, or senescent fibroblasts. These findings demonstrate that punctate dots containing LC3 do not always represent autophagic structures. Therefore, LC3 localization should be carefully interpreted, particularly if LC3 is overexpressed by transient transfection or if aggregates are formed within cells.
Kuma
Akiko
A
Department of Physiology and Cell Biology, Tokyo Medical and Dental University, Tokyo, Japan.
Matsui
Makoto
M
Mizushima
Noboru
N
D000818:Animals; D000957:Antigens, Viral, Tumor / Q000502:physiology; D016477:Artifacts; D001343:Autophagy / Q000502:physiology*; D015415:Biomarkers; D002461:Cell Line, Transformed; D002472:Cell Transformation, Viral; D002478:Cells, Cultured; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000378:metabolism; D007150:Immunohistochemistry; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008822:Mice, Transgenic; D008869:Microtubule-Associated Proteins / Q000378:metabolism*; D010714:Phosphatidylethanolamines / Q000378:metabolism; D011506:Proteins / Q000378:metabolism*; D013539:Simian virus 40 / Q000502:physiology; D014162:Transfection
D000957:Antigens, Viral, Tumor; D015415:Biomarkers; C480860:Map1lc3b protein, mouse; D008869:Microtubule-Associated Proteins; D010714:Phosphatidylethanolamines; D011506:Proteins; D049452:Green Fluorescent Proteins; C483858:phosphatidylethanolamine