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Related Concept Videos

Nuclear Export01:42

Nuclear Export

The nucleus restricts several proteins within and allows others to pass. The restricted proteins possess a nuclear retention sequence or NRS, anchoring them to the nuclear lamins and preventing their transport to the cytosol. The non-restricted proteins, after their synthesis, are transported to their site of action, such as the cytosol or other organelles, with the help of nuclear export signals or NES.
NES are of three types- the canonical 10-residue long leucine-rich signal and other...
Nuclear Protein Sorting01:34

Nuclear Protein Sorting

Nuclear protein sorting is the selective trafficking of histones, polymerases, gene regulatory proteins into the nucleus and exporting RNAs and ribosomes to the cytosol. It is a tightly controlled process that regulates gene expression within a cell.
Proteins targeted to the nucleus carry nuclear localization signals or NLS recognized by import receptors in the cytosol. Similarly, proteins with nuclear export signals are recognized by export receptors. Import and export receptors are...
Regulation of Nuclear Protein Sorting01:45

Regulation of Nuclear Protein Sorting

Nuclear protein sorting regulates nucleus composition and gene expression, crucial for determining the fate of a eukaryotic cell. Hence, the entry and exit of molecules across the nuclear envelope is a tightly controlled process. Nuclear protein sorting can be inhibited by one of the following ways: 1) masking cargo signal sequences, 2) modifying the nuclear receptor's affinity for cargo, 3) controlling the nuclear pore size, 4) retaining the cargo during its transit to the cytosol or the...
Phosphorylation01:02

Phosphorylation

The addition or removal of phosphate groups from proteins is the most common chemical modification that regulates cellular processes. These modifications can affect the structure, activity, stability, and localization of proteins within cells as well as their interactions with other proteins.
During phosphorylation, protein kinases transfer the terminal phosphate group of ATP to specific amino acid side chains of substrate proteins. Serine, threonine, and tyrosine are the most commonly...

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Updated: May 25, 2026

Identification of Cyclin-dependent Kinase 1 Specific Phosphorylation Sites by an In Vitro Kinase Assay
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Nucleofection induces transient eIF2α phosphorylation by GCN2 and PERK.

B R Anderson1, K Karikó, D Weissman

  • 1Department of Medicine, University of Pennsylvania, Philadelphia, PA 19104-6073, USA.

Gene Therapy
|February 4, 2012
PubMed
Summary
This summary is machine-generated.

Nucleofection, a method for delivering nucleic acids like mRNA, triggers rapid phosphorylation of eukaryotic initiation factor 2 alpha (eIF2α), reducing translation. This effect is mediated by GCN2 and PERK kinases, impacting nucleic acid-based therapies.

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Area of Science:

  • Molecular Biology
  • Cellular Biology
  • Biotechnology

Background:

  • Nucleofection is an efficient method for delivering nucleic acids (DNA, mRNA, siRNA) into challenging cell types.
  • Messenger RNA (mRNA) requires immediate translation post-delivery due to its rapid degradation.
  • Understanding translation dynamics after nucleofection is crucial for developing nucleic acid-based therapeutics.

Purpose of the Study:

  • To investigate the factors influencing translation following nucleofection.
  • To identify the kinases responsible for eukaryotic initiation factor 2 alpha (eIF2α) phosphorylation during nucleofection.
  • To assess the functional impact of eIF2α phosphorylation on translation.

Main Methods:

  • Investigated eIF2α phosphorylation kinetics post-nucleofection.
  • Assessed the role of GCN2, PERK, and PKR kinases in nucleofection-mediated eIF2α phosphorylation.
  • Measured translation efficiency following nucleofection.

Main Results:

  • Nucleofection induced rapid eIF2α phosphorylation independently of delivered nucleic acid.
  • This phosphorylation was dependent on GCN2 and PERK, but not PKR.
  • A significant reduction in translation was observed post-nucleofection due to eIF2α phosphorylation.

Conclusions:

  • Nucleofection activates GCN2 and PERK, leading to eIF2α phosphorylation and reduced translation.
  • These findings highlight the impact of nucleofection on the translational machinery.
  • Strategies to mitigate eIF2α phosphorylation are essential for advancing mRNA, DNA, and siRNA-based therapies.