cb616a277c2068eb27e3e9b764ac6749 pmc_10473220.pdf f1d3226d285e37ff562da62a6c3c942bb62bc343 pmc_10473220.pdf 3080229265966230cf9d71df28d109342d499d9435ccee4e846bb2d9dd586454 pmc_10473220.pdf Title: Arabidopsis metacaspase MC1 localizes in stress granules, clears protein aggregates, and delays senescence Subject: DOI: 10.1093/plcell/koad172; The Plant Cell, 35, 9, 2023-7-1.; Abstract: Stress granules (SGs) are highly conserved cytoplasmic condensates that assemble in response to stress and contribute to maintaining protein homeostasis. These membraneless organelles are dynamic, disassembling once the stress is no longer present. Persistence of SGs due to mutations or chronic stress has been often related to age-dependent protein-misfolding diseases in animals. Here, we find that the metacaspase MC1 is dynamically recruited into SGs upon proteotoxic stress in Arabidopsis (Arabidopsis thaliana). Two predicted disordered regions, the prodomain and the 360 loop, mediate MC1 recruitment to and release from SGs. Importantly, we show that MC1 has the capacity to clear toxic protein aggregates in vivo and in vitro, acting as a disaggregase. Finally, we demonstrate that overexpressing MC1 delays senescence and this phenotype is dependent on the presence of the 360 loop and an intact catalytic domain. Together, our data indicate that MC1 regulates senescence through its recruitment into SGs and this function could potentially be linked to its remarkable protein aggregate-clearing activity. Keywords: Author: Nerea Ruiz-Solaní Creator: Servigistics Arbortext Advanced Print Publisher 11.1.4546/W-x64 Producer: PDFlib+PDI 9.0.7p3 (C++/Win64) CreationDate: Wed Aug 16 18:11:42 2023 CEST ModDate: Tue Aug 29 09:51:41 2023 CEST Tagged: yes UserProperties: no Suspects: no Form: AcroForm JavaScript: no Pages: 20 Encrypted: no Page size: 595.276 x 782.362 pts Page rot: 0 File size: 6510364 bytes Optimized: yes PDF version: 1.6 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- NTAJIN+CronosPro-SemiboldIt Type 1C Custom yes yes yes 2074 0 BKIQDN+CronosPro-Semibold Type 1C Custom yes yes yes 2077 0 FONJYT+CronosPro-Regular Type 1C Custom yes yes yes 2080 0 JIMJCU+CronosPro-Bold Type 1C Custom yes yes yes 2083 0 XIPVTO+CronosPro-Italic Type 1C Custom yes yes yes 2086 0 FCIYJE+TimesNewRomanPSMT CID TrueType Identity-H yes yes yes 303 0 SEAYQQ+STIXGeneral-Regular Type 1C Custom yes yes yes 334 0 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ViewArea: CropBox ViewClip: CropBox PrintArea: CropBox PageClip: CropBox PageLayout: OneColumn PageMode: UseOutlines Language: en Outlines: Item: Title: Arabidopsis metacaspase MC1 localizes in stress granules, clears protein aggregates, and delays senescence Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@4671e53b Children: Item: Title: Introduction Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@2db7a79b Item: Title: Results Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@6950e31 Children: Item: Title: MC1 dynamically localizes to cytoplasmic SGs upon acute proteotoxic stress Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@b7dd107 Item: Title: The IDRs of MC1 are aggregation prone and confer insolubility in vitro Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@42eca56e Item: Title: The IDRs of MC1 regulate its dynamic recruitment into SGs Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@52f759d7 Item: Title: MC1 can specifically degrade aggregated proteins Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7cbd213e Item: Title: MC1 delays senescence Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@192d3247 Item: Title: Discussion Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@3ecd23d9 Children: Item: Title: MC1 as a SG component Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@569cfc36 Item: Title: MC1 can specifically clear protein aggregates Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@43bd930a Item: Title: Antiaging role of MC1: physiological implications of the role of MC1 in protein aggregate clearance Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@33723e30 Item: Title: Materials and methods Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@64f6106c Children: Item: Title: Materials used and growth conditions Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@553a3d88 Item: Title: Plasmid construction Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7a30d1e6 Item: Title: Protoplasts and plant transformation Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@5891e32e Item: Title: Yeast strains and spot dilution assays Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@cb0ed20 Item: Title: HEK293T cells transfection and protein extraction Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@8e24743 Item: Title: Microscopy analysis Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@74a10858 Item: Title: FRAP Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@23fe1d71 Item: Title: Protein purification Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@28ac3dc3 Item: Title: Enzymatic activity assays Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@32eebfca Item: Title: Preparation of TTR aggregates Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@4e718207 Item: Title: In vitro disaggregation assay Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1d371b2d Item: Title: Turbidity assay Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@543c6f6d Item: Title: Protein extraction and immunoblotting Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@13eb8acf Item: Title: Filter trap assay Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@51c8530f Item: Title: Heat treatments Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7403c468 Item: Title: Dark-induced senescence assay Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@43738a82 Item: Title: Chlorophyll analysis Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@c81cdd1 Item: Title: Pulse amplitude modulation (PAM) fluorometric measurements Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1fc2b765 Item: Title: RT-qPCR analysis Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@75881071 Item: Title: Bioinformatic analyses Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@2a70a3d8 Item: Title: Statistical analysis Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@289d1c02 Item: Title: Accession numbers Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@22eeefeb Item: Title: Acknowledgments Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@17d0685f Item: Title: Author Contributions Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@3891771e Item: Title: Supplemental data Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@78ac1102 Item: Title: Funding Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@2de8284b Item: Title: Data availability Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@396e2f39 Item: Title: References Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@a74868d Filters: FilterPipeline: FlateDecode Fonts: Type0: Font: BaseFont: GFOWWK+TimesNewRomanPS-ItalicMT Encoding: Identity-H ToUnicode: true Font: BaseFont: FCIYJE+TimesNewRomanPSMT Encoding: Identity-H ToUnicode: true Type1: Font: BaseFont: FONJYT+CronosPro-Regular FontSubset: true FirstChar: 1 LastChar: 102 FontDescriptor: FontName: FONJYT+CronosPro-Regular Flags: Nonsymbolic FontBBox: -179, -270, 1129, 886 FontFile3: true EncodingDictionary: BaseEncoding: WinAnsiEncoding Differences: true ToUnicode: true Font: BaseFont: JIMJCU+CronosPro-Bold FontSubset: true FirstChar: 1 LastChar: 86 FontDescriptor: FontName: JIMJCU+CronosPro-Bold Flags: Nonsymbolic, ForceBold FontBBox: -187, -295, 1137, 885 FontFile3: true EncodingDictionary: BaseEncoding: WinAnsiEncoding Differences: true ToUnicode: true Font: BaseFont: 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SEAYQQ+STIXGeneral-Regular Flags: Nonsymbolic FontBBox: -995, -455, 2000, 1055 FontFile3: true EncodingDictionary: BaseEncoding: WinAnsiEncoding Differences: true ToUnicode: true CIDFontType2: Font: BaseFont: FCIYJE+TimesNewRomanPSMT CIDSystemInfo: Registry: Adobe Registry: Identity Supplement: 0 FontDescriptor: FontName: FCIYJE+TimesNewRomanPSMT Flags: Nonsymbolic FontBBox: -568, -307, 2046, 1040 FontFile2: true Font: BaseFont: GFOWWK+TimesNewRomanPS-ItalicMT CIDSystemInfo: Registry: Adobe Registry: Identity Supplement: 0 FontDescriptor: FontName: GFOWWK+TimesNewRomanPS-ItalicMT Flags: Nonsymbolic, Italic FontBBox: -498, -307, 1333, 1023 FontFile2: true XMP: uuid:e5d496b7-3672-4ce5-a50e-d085e46d0f35 uuid:36AFB4CE-1E5E-47C3-2847-1C054D9989BD True PDFlib+PDI 9.0.7p3 (C++/Win64) application/pdf 10.1093/plcell/koad172 en Oxford University Press Arabidopsis metacaspase MC1 localizes in stress granules, clears protein aggregates, and delays senescence Nerea Ruiz-Solaní Jose Salguero-Linares Laia Armengot Jaime Santos Irantzu Pallarès Katarina P. van Midden Ujjal J. Phukkan Seda Koyuncu Júlia Borràs-Bisa Liang Li Crina Popa Frederik Eisele Anna Maria Eisele-Bürger Sandra Malgrem Hill Emilio Gutiérrez-Beltrán Thomas Nyström Marc Valls Ernesto Llamas David Vilchez Marina Klemenčič Salvador Ventura Nuria S. Coll DOI: 10.1093/plcell/koad172; The Plant Cell, 35, 9, 2023-7-1.; Abstract: Stress granules (SGs) are highly conserved cytoplasmic condensates that assemble in response to stress and contribute to maintaining protein homeostasis. These membraneless organelles are dynamic, disassembling once the stress is no longer present. Persistence of SGs due to mutations or chronic stress has been often related to age-dependent protein-misfolding diseases in animals. Here, we find that the metacaspase MC1 is dynamically recruited into SGs upon proteotoxic stress in Arabidopsis (Arabidopsis thaliana). Two predicted disordered regions, the prodomain and the 360 loop, mediate MC1 recruitment to and release from SGs. Importantly, we show that MC1 has the capacity to clear toxic protein aggregates in vivo and in vitro, acting as a disaggregase. Finally, we demonstrate that overexpressing MC1 delays senescence and this phenotype is dependent on the presence of the 360 loop and an intact catalytic domain. Together, our data indicate that MC1 regulates senescence through its recruitment into SGs and this function could potentially be linked to its remarkable protein aggregate-clearing activity. © The Author(s) 2023. Published by Oxford University Press on behalf of American Society of Plant Biologists. Journal The Plant Cell © The Author(s) 2023. Published by Oxford University Press on behalf of American Society of Plant Biologists. 1040-4651 1532-298X 35 9 3325 3344 10.1093/plcell/koad172 https://doi.org/10.1093/plcell/koad172 2023-08-16T21:41:42+05:30 2023-08-29T13:21:41+05:30 Servigistics Arbortext Advanced Print Publisher 11.1.4546/W-x64 2023-08-29T13:21:41+05:30 PStill version 1.84.42 1 PDF/UA Universal Accessibility Schema http://www.aiim.org/pdfua/ns/id/ pdfuaid part Integer internal Indicates, which part of ISO 14289 standard is followed Pages: Page: Sequence: 1 Annotations: Annotation: Subtype: Link Rect: 137, 544, 147, 554 Flags: Print Annotation: Subtype: Link Rect: 150, 547, 154, 559 Flags: Print Destination: 1990 Annotation: Subtype: Link Rect: 156, 547, 161, 559 Flags: Print Destination: 1990 Annotation: Subtype: Link Rect: 269, 544, 279, 554 Flags: Print Annotation: Subtype: Link Rect: 282, 547, 287, 559 Flags: Print Destination: 1990 Annotation: Subtype: Link Rect: 372, 544, 382, 554 Flags: Print Annotation: 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