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Nuclear Protein Sorting01:34

Nuclear Protein Sorting

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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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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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Directionality of Nuclear Transport01:42

Directionality of Nuclear Transport

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Ras-related nuclear protein or Ran is a small G protein that cycles between its GTP and GDP bound states. Ran specific regulators, a Ran GTPase Activating Protein or RanGAP present in the cytosol and a Ran guanine nucleotide exchange factor or RanGEF present inside the nucleus regulate GTP/GDP exchange. A high concentration of GTP inside the cells, in addition to this asymmetric distribution of  Ran-specific regulators, leads to a higher RanGTP concentration inside the nucleus. This...
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Rab Proteins01:14

Rab Proteins

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Rab proteins constitute the largest family of monomeric GTPases, of which 70 members are present in humans. Rab proteins and their effectors regulate consecutive stages of vesicle transport such as vesicle transport, docking, and fusion to the correct recipient membrane.
Rab proteins switch between a cytosolic, GDP-bound inactive state and a membrane-anchored, GTP-bound active state. By themselves, Rabs show slow rates of GDP/GTP exchange and GTP hydrolysis. Thus, Rab proteins are considered...
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RACE - Rapid Amplification of cDNA Ends02:35

RACE - Rapid Amplification of cDNA Ends

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Rapid Amplification of cDNA Ends, or RACE, is one of the most effective methods to obtain a full-length cDNA from an mRNA sequence between a known internal region to the unknown sequence at the 5’ or 3’ end. The unknown region is cloned in the cDNA by a gene-specific primer that binds the known end, and a hybrid primer that attaches a predefined anchor sequence to the unknown end of the cDNA. The sequence in between is amplified by PCR with an anchor primer and a gene-specific...
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A Rapid High-throughput Method for Mapping Ribonucleoproteins RNPs on Human pre-mRNA
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Bases estructurales para la disociación del complejo de importación nuclear por RanGTP

Soo Jae Lee1, Yoshiyuki Matsuura, Sai Man Liu

  • 1MRC Laboratory of Molecular Biology, Hills Rd, Cambridge CB2 2QH, UK.

Nature
|May 3, 2005
PubMed
Resumen
Este resumen es generado por máquina.

La importación de proteínas nucleares se basa en la importina-beta y el RanGTP. La unión de RanGTP a la importina-beta altera su estructura, liberando carga y regulando el transporte nuclear.

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Área de la Ciencia:

  • Biología Molecular Biología Molecular
  • Biología celular Biología celular.
  • Biología Estructural Biología estructural.

Sus antecedentes:

  • La importación de proteínas nucleares es esencial para la función celular.
  • La importina-beta actúa como un factor de transporte clave, a menudo utilizando la importina-alfa como un adaptador.
  • RanGTP impulsa el desmontaje de los complejos de importación dentro del núcleo.

Objetivo del estudio:

  • Para aclarar las bases estructurales del desmontaje de complejos de importación nuclear.
  • Para entender el mecanismo por el cual RanGTP libera carga de importin-beta.

Principales métodos:

  • Se utilizó la cristalografía de rayos X para determinar la estructura de la importina-beta de levadura (Kap95p) compleja con RanGTP.
  • Análisis de la estructura compleja para identificar las interfaces de interacción clave.

Principales resultados:

  • Se determinó la estructura de la importina-beta de levadura completa (Kap95p) compleja con RanGTP.
  • Se identificó un sitio crítico de interacción entre el bucle I del conmutador RanGTP y el arco carboxiterminal de Kap95p.
  • Esta interacción induce un cambio conformacional en Kap95p, impidiendo la unión de la carga.

Conclusiones:

  • La unión de RanGTP a la importina-beta causa un cambio conformacional alostérico.
  • Este mecanismo explica cómo RanGTP facilita la liberación de carga durante la importación nuclear.
  • Los hallazgos proporcionan una base estructural para la comprensión de la regulación del transporte nuclear.