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Nucleoporins redistribute inside the nucleus after cell cycle arrest induced by histone deacetylases inhibition.

Nucleus (Austin, Tex.) (2017-07-12)
Miguel Pérez-Garrastachu, Jon Arluzea, Ricardo Andrade, Alejandro Díez-Torre, Marta Urtizberea, Margarita Silió, Juan Aréchaga
ABSTRACT

Nucleoporins are the main components of the nuclear-pore complex (NPC) and were initially considered as mere structural elements embedded in the nuclear envelope, being responsible for nucleocytoplasmic transport. Nevertheless, several recent scientific reports have revealed that some nucleoporins participate in nuclear processes such as transcription, replication, DNA repair and chromosome segregation. Thus, the interaction of NPCs with chromatin could modulate the distribution of chromosome territories relying on the epigenetic state of DNA. In particular, the nuclear basket proteins Tpr and Nup153, and the FG-nucleoporin Nup98 seem to play key roles in all these novel functions. In this work, histone deacetylase inhibitors (HDACi) were used to induce a hyperacetylated state of chromatin and the behavior of the mentioned nucleoporins was studied. Our results show that, after HDACi treatment, Tpr, Nup153 and Nup98 are translocated from the nuclear pore toward the interior of the cell nucleus, accumulating as intranuclear nucleoporin clusters. These transitory structures are highly dynamic, and are mainly present in the population of cells arrested at the G0/G1 phase of the cell cycle. Our results indicate that the redistribution of these nucleoporins from the nuclear envelope to the nuclear interior may be implicated in the early events of cell cycle initialization, particularly during the G1 phase transition.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
L-Glutamine solution, 200 mM, solution, sterile-filtered, BioXtra, suitable for cell culture
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Penicillin-Streptomycin, with 10,000 units penicillin and 10 mg streptomycin per mL in 0.9% NaCl, 0.1 μm filtered, BioReagent, suitable for cell culture
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Monoclonal Anti-α-Tubulin antibody produced in mouse, ascites fluid, clone B-5-1-2
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SAHA, ≥98% (HPLC)
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Trichostatin A, Ready Made Solution, 5 mM in DMSO (0.2 μm-filtered), from Streptomyces sp.
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Anti-acetyl-Histone H3 Antibody, from rabbit
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Anti-Nup98 antibody, Rat monoclonal, clone 2H10, purified from hybridoma cell culture