TY - JOUR
T1 - Protrudin and PDZD8 contribute to neuronal integrity by promoting lipid extraction required for endosome maturation
AU - Shirane, Michiko
AU - Wada, Mariko
AU - Morita, Keiko
AU - Hayashi, Nahoki
AU - Kunimatsu, Reina
AU - Matsumoto, Yuki
AU - Matsuzaki, Fumiko
AU - Nakatsumi, Hirokazu
AU - Ohta, Keisuke
AU - Tamura, Yasushi
AU - Nakayama, Keiichi I.
N1 - Funding Information:
We thank S. Nagata and H. Sumimoto for providing the pEF-BOS-2×HA vector; T. Kitamura for providing the pMX-puro vector; M. Matsumoto for proteomics analysis; H. Takase for TEM analysis; K. Sawamoto for 3D construction of FIB-SEM images; T. Yasuda and A. Matsumoto for generation of CRISPR mutant mice; T. Ohnishi, Y. Kita, K. Matsuo, A. Kawajiri, R. Takemoto, I. Yamahata, H. Takahashi, Y. Ota, and H. Tashiro for technical assistance; T. Endo for discussion; and the Research Equipment Sharing Center at Nagoya City University for assistance. This work was supported in part by KAKENHI grants from Japan Society for the Promotion of Science (JSPS) and the Ministry of Education, Culture, Sports, Science, and Technology of Japan to M.S. (20H03255 and 20H04907) and to K.I.N. (18H05215).
Publisher Copyright:
© 2020, The Author(s).
PY - 2020/12/1
Y1 - 2020/12/1
N2 - Endosome maturation depends on membrane contact sites (MCSs) formed between endoplasmic reticulum (ER) and endolysosomes (LyLEs). The mechanism underlying lipid supply for this process and its pathophysiological relevance remains unclear, however. Here, we identify PDZD8—the mammalian ortholog of a yeast ERMES subunit—as a protein that interacts with protrudin, which is located at ER-LyLE MCSs. Protrudin and PDZD8 promote the formation of ER-LyLE MCSs, and PDZD8 shows the ability to extract various lipids from the ER. Overexpression of both protrudin and PDZD8 in HeLa cells, as well as their depletion in mouse primary neurons, impairs endosomal homeostasis by inducing the formation of abnormal large vacuoles reminiscent of those apparent in spastin- or REEP1-deficient neurons. The protrudin-PDZD8 system is also essential for the establishment of neuronal polarity. Our results suggest that protrudin and PDZD8 cooperatively promote endosome maturation by mediating ER-LyLE tethering and lipid extraction at MCSs, thereby maintaining neuronal polarity and integrity.
AB - Endosome maturation depends on membrane contact sites (MCSs) formed between endoplasmic reticulum (ER) and endolysosomes (LyLEs). The mechanism underlying lipid supply for this process and its pathophysiological relevance remains unclear, however. Here, we identify PDZD8—the mammalian ortholog of a yeast ERMES subunit—as a protein that interacts with protrudin, which is located at ER-LyLE MCSs. Protrudin and PDZD8 promote the formation of ER-LyLE MCSs, and PDZD8 shows the ability to extract various lipids from the ER. Overexpression of both protrudin and PDZD8 in HeLa cells, as well as their depletion in mouse primary neurons, impairs endosomal homeostasis by inducing the formation of abnormal large vacuoles reminiscent of those apparent in spastin- or REEP1-deficient neurons. The protrudin-PDZD8 system is also essential for the establishment of neuronal polarity. Our results suggest that protrudin and PDZD8 cooperatively promote endosome maturation by mediating ER-LyLE tethering and lipid extraction at MCSs, thereby maintaining neuronal polarity and integrity.
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U2 - 10.1038/s41467-020-18413-9
DO - 10.1038/s41467-020-18413-9
M3 - Article
C2 - 32917905
AN - SCOPUS:85090770132
VL - 11
JO - Nature Communications
JF - Nature Communications
SN - 2041-1723
IS - 1
M1 - 4576
ER -