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Spatial proteomics of ER tubules reveals CLMN, an ER-actin tether at focal adhesions that promotes cell migration

ABSTRACT: Ctrl = control; R = replicate; en = endogenously tagged; mNG = mNeonGreen LUM1_1034603: Ctrl_U2OS_R1 LUM1_1034604: Ctrl_U2OS_R2 LUM1_1034605 Ctrl_U2OS_R3 LUM1_1034613 U2OS_RTN4en_TurboID_mNG_R1 LUM1_1034614 U2OS_RTN4en_TurboID_mNG_R2 LUM1_1034615 U2OS_RTN4en_TurboID_mNG_R3 LUM1_1034621 U2OS_mNG_TurboID_CLIMP63en_R1 LUM1_1034622 U2OS_mNG_TurboID_CLIMP63en_R2 LUM1_1034623 U2OS_mNG_TurboID_CLIMP63en_R3 LUM1_1034628 U2OS_CANXen_TurboID_mNG_R1 LUM1_1034629 U2OS_CANXen_TurboID_mNG_R2 LUM1_1034630 U2OS_CANXen_TurboID_mNG_R3 LUM1_1034632 U2OS_mNG_TurboID_LBRen_R1 LUM1_1034633 U2OS_mNG_TurboID_LBRen_R2 LUM1_1034634 U2OS_mNG_TurboID_LBRen_R3 LUM1_1034636 U2OS_mNG_TurboID_Nesprin4_R1 LUM1_1034637 U2OS_mNG_TurboID_Nesprin4_R2 LUM1_1034638 U2OS_mNG_TurboID_Nesprin4_R3

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Homo Sapiens (ncbitaxon:9606)

SUBMITTER: W. Mike Henne

PROVIDER: MSV000096591 | MassIVE | Wed Dec 04 10:19:00 GMT 2024

REPOSITORIES: MassIVE

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Spatial proteomics of ER tubules reveals CLMN, an ER-actin tether at focal adhesions that promotes cell migration.

Project description:The endoplasmic reticulum (ER) is structurally and functionally diverse, yet how its functions are organized within morphological subdomains is incompletely understood. Utilizing TurboID-based proximity labeling and CRISPR knock-in technologies, here we map the proteomic landscape of the human ER and nuclear envelope. Spatial proteomics reveals enrichments of proteins into ER tubules, sheets, and nuclear envelope. We uncover an ER-enriched actin-binding protein, Calmin (CLMN), and define it as an ER-actin tether that localizes to focal adhesions adjacent to ER tubules. CLMN depletion perturbs focal adhesion disassembly, actin dynamics, and cell movement. Mechanistically, CLMN-depleted cells also exhibit defects in calcium signaling near ER-actin interfaces, suggesting CLMN promotes calcium signaling near adhesions to facilitate their disassembly. Collectively, we map the sub-organelle proteome landscape of the ER, identify CLMN as an ER-actin tether, and describe a non-canonical mechanism by which ER tubules engage actin to regulate cell migration.

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Spatial regulation of MCAK promotes cell polarization and focal adhesion turnover to drive robust cell migration.

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2023-06-26 | GSE205966 | GEO

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