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In eukaryotic cells, nascent mRNA transcripts need to undergo many post-transcriptional modifications to reach the cell cytoplasm and translate into functional proteins. For a long time, transcription and pre-mRNA processing were considered two independent events that occur sequentially in the cell. However, it has now been well established that transcription and pre-mRNA processing are two simultaneous processes that are precisely regulated inside the cell.
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Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
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Regulated mRNA Transport02:22

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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 Export of mRNA02:31

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Before mRNAs are exported to the cytoplasm, it is crucial to check each mRNA for structural and functional integrity. Eukaryotic cells use several different mechanisms, collectively known as mRNA surveillance, to look for irregularities in mRNAs. Irregular or aberrant mRNA are rapidly degraded by various enzymes. If a defective mRNA escapes the surveillance, it would be translated into a protein which would either be non-functional or not function properly. One of the primary irregularities in...
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In eukaryotic cells, transcripts made by RNA polymerase are modified and processed before exiting the nucleus. Unprocessed RNA is called precursor mRNA or pre-mRNA to distinguish it from mature mRNA.
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The structure and stability of mRNA molecules regulates gene expression, as mRNAs are a key step in the pathway from gene to protein. In eukaryotes, the half-life of mRNA varies from a few minutes up to several days. mRNA stability is essential in growth and development. The absence of the proteins regulating its stability, such as tristetraprolin in mice, can cause systemic issues, including bone marrow overgrowth, inflammation, and autoimmunity.
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Separation of Rat Epidermis and Dermis with Thermolysin to Detect Site-Specific Inflammatory mRNA and Protein
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La estructura del ARNm determina la especificidad de una separación de fase impulsada por el poliQ

Erin M Langdon1, Yupeng Qiu2, Amirhossein Ghanbari Niaki2

  • 1Department of Biology, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA.

Science (New York, N.Y.)
|April 14, 2018
PubMed
Resumen

Los ARN mensajeros (ARNm) usan su estructura para autoasociarse, formando compartimentos celulares distintos. Las interacciones de las proteínas mantienen estos compartimentos líquidos impulsados por ARN, creando diversidad celular.

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Área de la Ciencia:

  • Biología celular
  • Biología molecular
  • La bioquímica

Sus antecedentes:

  • Los compartimentos sin membrana celular se forman a través de la separación de fase líquido-líquido (LLPS).
  • Los mecanismos que establecen composiciones moleculares distintas dentro de estos compartimentos ricos en ARN siguen sin estar claros.

Objetivo del estudio:

  • Investigar cómo los ARN mensajeros (ARNm) contribuyen a la formación y mantenimiento de compartimentos líquidos distintos.
  • Determinar el papel de la estructura secundaria de ARN en el reclutamiento molecular y la exclusión de estos compartimentos.

Principales métodos:

  • Se estudió la autoasociación del ARNm impulsada por la estructura secundaria del ARN.
  • Se ha investigado la influencia de la proteína Whi3 en la estructura y dinámica del ARN.
  • Se analizaron las fluctuaciones moleculares dependientes de la secuencia de ARN dentro de los compartimentos.

Principales resultados:

  • La estructura secundaria del ARN permite la autoasociación del ARNm, lo que dicta la pertenencia al compartimento.
  • La proteína Whi3 induce cambios conformacionales en el ARN, modulando las fluctuaciones moleculares.
  • Las interacciones ARN-ARN basadas en la estructura impulsan el ensamblaje de gotas distintas y el mantenimiento de la identidad.

Conclusiones:

  • La forma y la estructura del ARN son críticas para la formación de diversos compartimentos líquidos ricos en ARN que coexisten.
  • Dinámica conformacional mediada por proteínas del compartimiento de estabilización del ARN.
  • Esto proporciona un marco para comprender la heterogeneidad del compartimento sin membrana celular.