Supplemental Material for Germoglio and Adamo, 2018

2018-06-08T12:48:18Z (GMT) by Germoglio M. Adamo A.
<p>The evolutionary conserved RAD-51 protein is essential for homologous recombination in the germ line as well as homologous repair of DNA double strand breaks in all eukaryotic cells. In the nematode <i>Caenorhabditis elegans</i> the <i>rad-51</i> gene is transcribed into messenger-RNAs potentially coding three alternative protein isoforms. Null <i>rad-51</i> alleles display embryonic lethality, severe defects in chromosome structure and high level of germ line apoptosis. To dissect its functions, we genetically modified the <i>C. elegans rad-51</i> gene by CRISPR/Cas9 genome-editing technology, obtaining a separation-of-function (sfi-) mutant allele that only disrupts the long transcript isoform. This mutant shows no defects in an otherwise wild-type meiosis and is able to activate physiological germ cell death, which occurs at the late pachytene stage. However, although the mutant is competent in DNA damage checkpoint activation after exposure to ionizing radiation, it is defective for induction of DNA-damage induced apoptosis in meiotic germ cells. These results suggest that RAD-51 plays a novel role in germline apoptosis independent of RAD-51-mediated strand invasion for homologous recombination.</p>