Role of RadA and DNA Polymerases in Recombination-Associated DNA Synthesis in Hyperthermophilic Archaea.

Biomolecules
Gaëlle HogrelDidier Flament

Abstract

Among the three domains of life, the process of homologous recombination (HR) plays a central role in the repair of double-strand DNA breaks and the restart of stalled replication forks. Curiously, main protein actors involved in the HR process appear to be essential for hyperthermophilic Archaea raising interesting questions about the role of HR in replication and repair strategies of those Archaea living in extreme conditions. One key actor of this process is the recombinase RadA, which allows the homologous strand search and provides a DNA substrate required for following DNA synthesis and restoring genetic information. DNA polymerase operation after the strand exchange step is unclear in Archaea. Working with Pyrococcus abyssi proteins, here we show that both DNA polymerases, family-B polymerase (PolB) and family-D polymerase (PolD), can take charge of processing the RadA-mediated recombination intermediates. Our results also indicate that PolD is far less efficient, as compared with PolB, to extend the invaded DNA at the displacement-loop (D-loop) substrate. These observations coincide with previous genetic analyses obtained on Thermococcus species showing that PolB is mainly involved in DNA repair without being essential ...Continue Reading

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Citations

Dec 2, 2020·Open Biology·Patricia Pérez-ArnaizThorsten Allers
Oct 30, 2020·Biomolecules·Craig J Marshall, Thomas J Santangelo
Jun 12, 2021·Frontiers in Microbiology·Florence GarnierMarc Nadal

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Methods Mentioned

BETA
gel filtration
protein assay
electrophoresis
electrophoretic mobility shift

Software Mentioned

ImageQuant
ProtParam
ExPASy
Genecust

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