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Homologous Recombination02:31

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The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...
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Fixing Double-strand Breaks02:04

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The double-stranded structure of DNA has two major advantages. First, it serves as a safe repository of genetic information where one strand serves as the back-up in case the other strand is damaged. Second, the double-helical structure can be wrapped around proteins called histones to form nucleosomes, which can then be tightly wound to form chromosomes. This way, DNA chains up to 2 inches long can be contained within microscopic structures in a cell. A double-stranded break not only damages...
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Translesion DNA Polymerases02:10

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Translesion (TLS) polymerases rescue stalled DNA polymerases at sites of damaged bases by replacing the replicative polymerase and installing a nucleotide across the damaged site. Doing so, TLS allows additional time for the cell to repair the damage before resuming regular DNA replication.
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LTR Retrotransposons03:08

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LTR retrotransposons are class I transposable elements with long terminal repeats flanking an internal coding region. These elements are less abundant in mammals compared to other class I transposable elements. About 8 percent of human genomic DNA comprises LTR retrotransposons. Some of the common examples of LTR retrotransposons are Ty elements in yeast and Copia elements in Drosophila.
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Other than maintaining genome stability via DNA repair, homologous recombination plays an important role in diversifying the genome. In fact, the recombination of sequences forms the molecular basis of genomic evolution. Random and non-random permutations of genomic sequences create a library of new amalgamated sequences. These newly formed genomes can determine the fitness and survival of cells. In bacteria, homologous and non-homologous types of recombination lead to the evolution of new...
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Función de las transcriptasas inversas similares al intrón del grupo II en la reparación de la rotura de doble cadena

Seung Kuk Park1, Georg Mohr1, Jun Yao1

  • 1Departments of Molecular Biosciences and Oncology, University of Texas at Austin, Austin, TX 78712, USA.

Cell
|September 16, 2022
PubMed
Resumen

Las bacterias poseen transcriptasas inversas (RT) que funcionan en la reparación del ADN. Estas enzimas realizan la síntesis de ADN de traslación y la reparación de la ruptura de doble cadena (DSBR) a través de la unión final mediada por microhomología (MMEJ).

Palabras clave:
Alt-EJLa polimerasa de reparación del ADNunión de extremo alternativosecuenciación de alto rendimientoelemento R2 de los insectostranscriptasa inversa no retroviralobjetivotranscriptasa inversa intrónica del grupo II térmicamente estable

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Área de la Ciencia:

  • Biología molecular
  • La genética
  • La bioquímica

Sus antecedentes:

  • Las bacterias codifican las transcriptasas inversas (RT) estructuralmente relacionadas con las RT intrónicas del grupo II, con funciones en gran medida desconocidas.
  • Los RT de intrones del grupo II son elementos genéticos móviles conocidos involucrados en el empalme y la retrohomología del ARN.

Objetivo del estudio:

  • Investigar la función de un RT similar al intrón del grupo II (G2L4 RT) de Pseudomonas aeruginosa.
  • Determinar si las RT bacterianas pueden funcionar en las vías de reparación del ADN, específicamente en la síntesis de translesión del ADN y en la reparación de la ruptura de doble hebra (DSBR).

Principales métodos:

  • Caracterización bioquímica del G2L4 RT.
  • Expresión de G2L4 RT en Escherichia coli.
  • Mutagénesis dirigida al sitio para analizar los residuos activos del sitio (YIDD vs. YADD).
  • Análisis comparativo con la polimerasa theta de reparación del ADN humano y otras RT de retroelementos no LTR.

Principales resultados:

  • G2L4 RT, con un sitio activo YIDD, exhibe actividades de reparación del ADN en su huésped nativo y en E. coli.
  • G2L4 RT realiza la síntesis de ADN de traslación y DSBR a través de la unión final mediada por microhomología (MMEJ).
  • Sus actividades bioquímicas son similares a las de la polimerasa theta de reparación del ADN humano.
  • Las sustituciones recíprocas del sitio activo revelaron que la isoleucina favorece la MMEJ y la alanina favorece la extensión del primer tanto en la RT intrónica del grupo II como en la RT G2L4.

Conclusiones:

  • Los RT similares a los intrones del grupo II bacteriano poseen capacidades inherentes de reparación del ADN, incluida la DSBR.
  • Estas funciones se basan en características estructurales conservadas compartidas con los RT retroelementos no LTR eucariotas.
  • Los RT bacterianos pueden desempeñar un papel conservado en DSBR en diversos organismos.