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The nuclear permeability barrier depends on closure of nuclear envelope (NE) holes. Here, we investigate closure of the NE opening surrounding the meiotic spindle in C. elegans oocytes. ESCRT-III components accumulate at the opening but are not required for nuclear closure on their own. 3D analysis revealed cytoplasmic membranes directly adjacent to NE holes containing meiotic spindle microtubules. We demonstrate that the NE protein phosphatase, CNEP-1/CTDNEP1, controls de novo glycerolipid synthesis through lipin to prevent invasion of excess ER membranes into NE holes and a defective NE permeability barrier. Loss of NE adaptors for ESCRT-III exacerbates ER invasion and nuclear permeability defects in cnep-1 mutants, suggesting that ESCRTs restrict excess ER membranes during NE closure. Restoring glycerolipid synthesis in embryos deleted for CNEP-1 and ESCRT components rescued NE permeability defects. Thus, regulating the production and feeding of ER membranes into NE holes together with ESCRT-mediated remodeling is required for nuclear closure.
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Nuclear envelope assembly relies on CHMP-7 in the absence of BAF-LEM-mediated hole closure
Barrier-to-autointegration (BAF) is a DNA binding protein that crosslinks chromatin to allow mitotic nuclear envelope (NE) assembly. The Lap2b-Emerin-Man1(LEM)-domain protein LEMD2 and ESCRTII/III hybrid protein CHMP7 close NE holes surrounding spindle microtubules (MTs). BAF binds LEM-domain family proteins, which repairs NE ruptures in interphase, but whether BAF-LEM binding participates in NE hole closure around spindle MTs is not known. Here, we take advantage of the stereotypical event of NE formation in fertilized C. elegans oocytes to show that BAF-LEM binding and LEM-2LEMD2-CHMP-7 have distinct roles in NE closure around spindle MTs. LEM-2/EMR-1emerin function redundantly with BAF-1 in NE closure. Compromising BAF-LEM binding revealed an additional role for EMR-1emerin in maintenance of the NE permeability barrier. In the absence of BAF-LEM binding, LEM-2-CHMP-7 are required for NE assembly and embryo survival. The winged helix domain of LEM-2 recruits CHMP-7 to the NE in C. elegans and a LEM-2-independent nucleoplasmic pool of CHMP-7 also contributes to NE stability. Thus, NE hole closure surrounding spindle MTs requires redundant mechanisms that safeguard against failure in NE assembly to support embryogenesis.
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- Award ID(s):
- 1846010
- PAR ID:
- 10467806
- Publisher / Repository:
- The Company of Biologists
- Date Published:
- Journal Name:
- Journal of Cell Science
- ISSN:
- 0021-9533
- Format(s):
- Medium: X
- Sponsoring Org:
- National Science Foundation
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