Skip to Content
Merck
  • Fragile X-Related Protein 1 Regulates Nucleoporin Localization in a Cell Cycle-Dependent Manner.

Fragile X-Related Protein 1 Regulates Nucleoporin Localization in a Cell Cycle-Dependent Manner.

Frontiers in cell and developmental biology (2022-01-04)
Arantxa Agote-Arán, Junyan Lin, Izabela Sumara
ABSTRACT

Nuclear pore complexes (NPCs) are embedded in the nuclear envelope (NE) where they ensure the transport of macromolecules between the nucleus and the cytoplasm. NPCs are built from nucleoporins (Nups) through a sequential assembly order taking place at two different stages during the cell cycle of mammalian cells: at the end of mitosis and during interphase. In addition, fragile X-related proteins (FXRPs) can interact with several cytoplasmic Nups and facilitate their localization to the NE during interphase likely through a microtubule-dependent mechanism. In the absence of FXRPs or microtubule-based transport, Nups aberrantly localize to the cytoplasm forming the so-called cytoplasmic nucleoporin granules (CNGs), compromising NPCs' function on protein export. However, it remains unknown if Nup synthesis or degradation mechanisms are linked to the FXRP-Nup pathway and if and how the action of FXRPs on Nups is coordinated with the cell cycle progression. Here, we show that Nup localization defects observed in the absence of FXR1 are independent of active protein translation. CNGs are cleared in an autophagy- and proteasome-independent manner, and their presence is restricted to the early G1 phase of the cell cycle. Our results thus suggest that a pool of cytoplasmic Nups exists that contributes to the NPC assembly specifically during early G1 to ensure NPC homeostasis at a short transition from mitosis to the onset of interphase.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Anti-FXR1 antibody produced in rabbit, Prestige Antibodies® Powered by Atlas Antibodies, affinity isolated antibody, buffered aqueous glycerol solution
Sigma-Aldrich
Pepstatin A, microbial, ≥90% (HPLC)
Sigma-Aldrich
E-64d, protease inhibitor
Sigma-Aldrich
Z-Leu-Leu-Leu-al, ≥90% (HPLC)
Sigma-Aldrich
Cycloheximide solution, Ready-Made Solution, microbial, 100 mg/mL in DMSO, Suitable for cell culture
Sigma-Aldrich
RIPAb+ FXR1 - RIP Validated Antibody and Primer Set, from mouse
Sigma-Aldrich
Cdk1 Inhibitor IV, RO-3306, RO-3306 is a cell-permeable, potent and ATP-competitive inhibitor of Cdk1 (Ki = 35 nM and 110 nM for Cdk1/B1 and Cdk1/A, respectively).