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Related Concept Videos

Nuclear Localization Signals and Import01:46

Nuclear Localization Signals and Import

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Proteins targeted to the nucleus carry short stretches of amino acid sequences called the nuclear localization signal or NLS. Classical nuclear localization signals are of two types: monopartite and bipartite NLS. Monopartite classical NLS (cNLS) consists of a single cluster of 4-8 amino acids. Bipartite cNLS consists of two clusters of  2-3 amino acids and a 9-12 residue long proline-rich linker bridging the two clusters. Signal clusters are rich in positively charged amino acids such as...
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Nuclear Export01:42

Nuclear Export

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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...
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Regulation of Nuclear Protein Sorting01:45

Regulation of Nuclear Protein Sorting

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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...
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Regulated mRNA Transport02:22

Regulated mRNA Transport

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In eukaryotes, transcription and translation are compartmentalized; an mRNA is first synthesized in the nucleus and then selectively transported to the cytoplasm for protein synthesis. Before transport, a pre-mRNA undergoes several steps of post-transcriptional modifications including splicing, 5' capping, and the addition of a poly-adenine tail. Various proteins bind to the pre-mRNA during these modifications. The mRNA transport takes place with the help of multiple proteins playing...
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Nuclear Protein Sorting01:34

Nuclear Protein Sorting

4.5K
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...
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Mitochondrial Precursor Proteins01:39

Mitochondrial Precursor Proteins

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Mitochondrial precursors are partially unfolded or loosely folded polypeptide chains. Newly synthesized precursors are inhibited from spontaneously folding into their native conformation by the cytosolic chaperones, heat shock proteins 70 (Hsp70), and mitochondrial import stimulation factors (MSFs). Precursors bound to MSFs are guided to the TOM70-TOM37 receptors, while precursors bound to Hsp70  chaperones are targetted to TOM20-TOM22 receptor complexes.
Most of the mitochondrial...
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Related Experiment Video

Updated: Jun 6, 2025

Heterokaryon Technique for Analysis of Cell Type-specific Localization
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Sequence specificity of an essential nuclear localization sequence in Mcm3.

Ziyi Wang1,2, Yun Jing Zhang2, Qian-Yi Zhang1

  • 1Biomedical Science graduate program, School of Medicine, University of California at San Diego.

Biorxiv : the Preprint Server for Biology
|November 28, 2024
PubMed
Summary

The precise positioning of basic amino acids in Mcm3's nuclear localization sequence (NLS) is crucial for importin interaction and nuclear import. Disrupting this impacts DNA replication and cell growth.

Keywords:
AlphaFold 3MCM loadingNuclear localization sequence (NLS)importin alpha

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Area of Science:

  • Cell Biology
  • Molecular Biology
  • Structural Biology

Background:

  • Nuclear localization sequences (NLSs) mediate protein import into the cell nucleus.
  • Identifying NLSs is challenging due to sequence variability.
  • Mcm3 is a vital subunit of the replicative DNA helicase.

Purpose of the Study:

  • To characterize the sequence specificity of the Mcm3 NLS.
  • To understand the structural basis of Mcm3 nuclear import.
  • To develop a framework for analyzing monopartite NLSs.

Main Methods:

  • Site-directed mutagenesis of the Mcm3 NLS.
  • AlphaFold 3 (AF3) protein structure modeling.
  • Analysis of protein-nuclear import and chromatin loading.

Main Results:

  • The precise arrangement of basic residues in the Mcm3 NLS dictates importin binding.
  • Mutations disrupting NLS-importin interactions impair Mcm3 nuclear import.
  • Impaired nuclear import of Mcm3 leads to defective MCM chromatin loading and reduced cell growth.

Conclusions:

  • The study provides a structure-guided method for predicting monopartite NLSs.
  • Key characteristics of the Mcm3 NLS are shared with other NLSs, like SV40 large T antigen.
  • Understanding NLS sequence specificity is critical for nuclear transport regulation.