Identification of nuclear pore proteins at plasmodesmata

Abstract

Here we used bioinformatics, proteomics and fluorescence imaging to identify proteins with similarities to phase separating FG-NUPs at PD. We identified 20 NUPs in PD fractions, and validated dual localization to NPC and PD for 7 NUPs. Structured illumination microscopy detected the transmembrane anchor NUP CPR5 at both orifices of PD. cpr5 mutants showed reduced intercellular transport of SHR from stele to endodermis. The identification of FG-NUPs at PD is consistent with the recruitment of NUPs in the green lineage to form a PD pore gating complex consistent with liquid phase separation domains as diffusion barriers at PD.

Preprint

A bioRxiv preprint of the associated publication is available at https://doi.org/10.1101/2024.09.02.610746 and here

Raw data and metadata can be found in "assays"

• NUP localization
  • transiently expressed
  • stably expressed
• topology of CPR5
• structured illumination microscopy (SIM) transiently expressing CPR5 and PDLP5
• transport assays
  • bombardment
  • DropANdSee (DANS)
  • short root SHR-GFP
• sequencing results for cpr5G420D

Contributors

• Till Moritz Schladt
  • email: till.schladt@hhu.de
  • Affiliation: Heinrich Heine Univeryity, Düsseldorf
  • ORCID: 0000-0002-9160-4664
• Miras Manuel
  • Affiliation: CEBAS-CSIC, Heinrich-Heine-Universität Düsseldorf Mathematisch-Naturwissenschaftliche Fakultät, Insitute of Molecular Physiology Heinrich-Heine-Universität Düsseldorf Mathematisch-Naturwissenschaftliche Fakultät, Institute Molecular Physiology;Abiopep SL, ;Centro de Edafología y Biología Aplicada del Segura, Plant Virology;Centro de Edafología y Biología Aplicada del Segura, Plant Virology
  • ORCID: 0000-0002-3636-0265
• Jona Obinna Ejike
  • Affiliation: Heinrich Heine Universität, Düsseldorf
  • ORCID: 0000-0002-4601-8551
• Mathieu Pottier
  • Affiliation: Université de Lorraine, UMR 1136 Université de Lorraine/INRAE Interactions Arbres Microorganismes (IAM);Heinrich-Heine-Universität Düsseldorf, Institute for Molecular Physiology;University of Liège, Integrative Biological Sciences (InBioS);Université catholique de Louvain, Louvain Institute of Biomolecular Science and Technology (LIBST);CNRS Délégation Ile-de-France Sud, Institut des Sciences du Végétal (ISV)
  • ORCID: 0000-0003-1551-4699
• Lin Xi
  • Affiliation: University of Hohenheim
  • ORCID: 0000-0002-6811-9505
• Andrea Restrepo
  • Affiliation: Heinrich-Heine-Universität Düsseldorf, Institute for Molecular Physiology
  • ORCID: 0000-0003-2870-394X
• Masayoshi Nakamura
  • Affiliation: Nagoya University, Institute of Transformative Bio-Molecules;Nagoya University, Institute of Transformative Bio-Molecules;Carnegie Institution for Science, Department of Plant Biology;Nara Institute of Science and Technology, Department of Biological Science
  • ORCID: 0000-0002-7107-4942
• Niklas Pütz
  • Affiliation: Heinrich Heine Universität, Düsseldorf
  • ORCID: 0009-0006-1153-5918
• Sebastian Hänsch
  • Affiliation: Heinrich Heine University Düsseldorf, Center for Advanced Imaging;Heinrich Heine University Düsseldorf, Center for Advanced Imaging
  • ORCID: 0000-0002-7762-2516
• Chen Gao
  • Affiliation: University of Copenhagen, Department of Plant and Environmental Sciences
  • ORCID: 0000-0003-4750-3900
• Julia Engelhorn
  • Affiliation: University of Montpellier, IRD;Heinrich Heine University Düsseldorf, Institute for Molecular Physiology;University of Warwick, School of Life Sciences;CEA Grenoble, Cell & Plant Physiology Laboratory
  • ORCID: 0000-0002-6680-3012
• Marcel Dickmanns
  • Affiliation: Heinrich-Heine-Universität Düsseldorf, ;Max Planck Institute of Biochemistry,
  • ORCID: 0000-0003-4001-707X
• Gwendolyn Davis
  • Affiliation: University of Warwick, Life Sciences;University of Cambridge, Plant Sciences
  • ORCID: 0000-0003-4704-4863
• Ahan Dalal
  • Affiliation: Hebrew University of Jerusalem, Plant Science;Tel Aviv University, Plant Science
  • ORCID: 0000-0003-1876-1484
• Sven Gombos
  • Affiliation: University of Hohenheim, Plant Systems Biology
  • ORCID: 0000-0003-4177-526X
• Ronja Lange
  • Affiliation: Heinrich Heine Universität, Düsseldorf
  • ORCID: 0009-0009-2475-2438
• Rüdiger Simon
  • Affiliation: Heinrich Heine University, Biology
  • ORCID: 0000-0002-1317-7716
• Waltraud Schulze
  • Affiliation: University of Hohenheim
  • ORCID: 0000-0001-9957-7245
• Wolf B. Frommer
  • Affiliation: Heinrich-Heine-Universitat Düsseldorf, Institute for Molecular Physiology;ITbM, ;Carnegie Institution for Science, Plant Biology;Stanford University, Biology;Carnegie Institution for Science, Plant Biology;Stanford University, Biology;Carnegie Institution for Science, Plant Biology;University of Tübingen, Biology
  • ORCID: 0000-0001-6465-0115