Article ; Online: Membrane contact site detection (MCS-DETECT) reveals dual control of rough mitochondria-ER contacts.
2023 Volume 223, Issue 1
Abstract: Identification and morphological analysis of mitochondria-ER contacts (MERCs) by fluorescent microscopy is limited by subpixel resolution interorganelle distances. Here, the membrane contact site (MCS) detection algorithm, MCS-DETECT, reconstructs ... ...
| Abstract | Identification and morphological analysis of mitochondria-ER contacts (MERCs) by fluorescent microscopy is limited by subpixel resolution interorganelle distances. Here, the membrane contact site (MCS) detection algorithm, MCS-DETECT, reconstructs subpixel resolution MERCs from 3D super-resolution image volumes. MCS-DETECT shows that elongated ribosome-studded riboMERCs, present in HT-1080 but not COS-7 cells, are morphologically distinct from smaller smooth contacts and larger contacts induced by mitochondria-ER linker expression in COS-7 cells. RiboMERC formation is associated with increased mitochondrial potential, reduced in Gp78 knockout HT-1080 cells and induced by Gp78 ubiquitin ligase activity in COS-7 and HeLa cells. Knockdown of riboMERC tether RRBP1 eliminates riboMERCs in both wild-type and Gp78 knockout HT-1080 cells. By MCS-DETECT, Gp78-dependent riboMERCs present complex tubular shapes that intercalate between and contact multiple mitochondria. MCS-DETECT of 3D whole-cell super-resolution image volumes, therefore, identifies novel dual control of tubular riboMERCs, whose formation is dependent on RRBP1 and size modulated by Gp78 E3 ubiquitin ligase activity. |
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| MeSH term(s) | Humans ; Endoplasmic Reticulum/metabolism ; HeLa Cells ; Mitochondria/metabolism ; Mitochondrial Membranes/metabolism ; Ubiquitin-Protein Ligases/metabolism ; Ubiquitination ; COS Cells ; Animals ; Chlorocebus aethiops ; Ribosomes/metabolism | |||||
| Chemical Substances | Ubiquitin-Protein Ligases (EC 2.3.2.27) | |||||
| Language | English | |||||
| Publishing date | 2023-11-10 | |||||
| Publishing country | United States | |||||
| Document type | Journal Article ; Research Support, Non-U.S. Gov't | |||||
| ZDB-ID | 218154-x | |||||
| ISSN | 1540-8140 ; 0021-9525 | |||||
| ISSN (online) | 1540-8140 | |||||
| ISSN | 0021-9525 | |||||
| DOI | 10.1083/jcb.202206109 | |||||
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| Database | MEDical Literature Analysis and Retrieval System OnLINE |
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