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Series GSE178665 Query DataSets for GSE178665
Status Public on Mar 28, 2022
Title ESCRT-I controls lysosomal membrane protein homeostasis and restricts MCOLN1-dependent TFEB/TFE3 signaling.
Organism Homo sapiens
Experiment type Expression profiling by high throughput sequencing
Summary Within the endolysosomal pathway in mammalian cells, ESCRT complexes facilitate degradation of membrane proteins from limiting membranes of late endosomes. Recent studies revealed that yeast ESCRT proteins also sort for degradation, ubiquitinated proteins from vacuolar membrane. However, whether mammalian ESCRTs perform similar function at lysosomes remained unknown. Here, we studied the involvement of mammalian ESCRT-I in maintaining lysosomal membrane homeostasis and its implication in lysosome-related signaling. We show that ESCRT-I restricts the size of lysosomes and promotes degradation of lysosomal Ca2+ channel, MCOLN1 protein. ESCRT-I depletion induced transcriptional response related to MiT-TFE signaling and cholesterol biosynthesis, pointing to lysosomal dysfunction. The lack of ESCRT-I promoted abnormal cholesterol accumulation on lysosomes and activated TFEB/TFE3 transcription factors in Ca2+-MCOLN1-dependent, but lipid-independent manner. Hence, our study provides evidence that ESCRT-I maintains lysosomal homeostasis and elucidates MiT-TFE regulatory mechanism activated in response to ESCRT-I deprivation
Overall design The aim of the study was to analyze by RNA sequencing the transcriptional changes upon single siRNA-mediated depletion of two core component of ESCRT-I complex, Tsg101 or Vps28 subunit. As a model, human colorectal RKO cell line was used. 3 biological replicates were prepared for each experimental condition. The conditions were: 1/RKO transfected with non -targeting siRNA (siCtrl#1), 2/ RKO transfected with non-targeting siRNA (siCtrl#2), 3/ RKO transfected with siRNA targeting TSG101, sequence #1 (siTsg101#1), 4/ RKO transfected with siRNA targeting TSG101, sequence #2 (siTsg101#2), 5/ RKO transfected with siRNA targeting VPS28, sequence #1 (siVps28#1), 6/ RKO transfected with siRNA targeting VPS28, sequence #2 (siVps28#2). siCtrl#1-transfected cells served as the reference sample.
Contributor(s) Wróbel M, Szymańska E, Budick-Harmelin N, Kolmus K, Goryca K, Dąbrowska M, Paziewska A, Mikula M, Cendrowski J, Miączyńska M
Citation(s) 35354596
Submission date Jun 22, 2021
Last update date Apr 20, 2022
Contact name Michał Mikula
Phone +48225462655
Organization name Maria Skłodowska-Curie National Research Institute of Oncology
Department Genetics
Street address Roentgena 5
City Warsaw
ZIP/Postal code 02781
Country Poland
Platforms (1)
GPL17303 Ion Torrent Proton (Homo sapiens)
Samples (18)
GSM5395653 RKO_siCtrl#1_1
GSM5395654 RKO_siCtrl#2_1
GSM5395655 RKO_siTsg101#1_1
BioProject PRJNA739978
SRA SRP325061

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Supplementary file Size Download File type/resource
GSE178665_RAW.tar 1.3 Mb (http)(custom) TAR (of TXT)
SRA Run SelectorHelp
Raw data are available in SRA
Processed data provided as supplementary file

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