Poly (ADP-ribose) polymerases (PARPs) are specific enzymes catalyzing the post-translational addition of ADP-ribose moieties to proteins. In most eukaryotic cells, their primary protein targets are involved in DNA recombination, repair, and chromosome maintenance. Even though this group of enzymes is quite common in both eukaryotes and prokaryotes, no PARP homologs have been described so far in ascomycetous yeasts, leaving their potential roles in this group of organisms unexplored. Analysis of transcriptome of telomerase-deficient cells of the yeast Yarrowia lipolytica resulted in a serendipitous identification of the gene PYL1 encoding the first candidate of PARP homolog in yeasts. Here, we show that Pyl1 protein is a functional PARP that undergoes auto-PARylation and we identified several of its protein targets in vivo. The fact that the major targets of PARylation are both subunits of the Ku70/Ku80 complex and that overexpression of PYL1 leads to elongation of telomeres support the role of Pyl1 in the adaptation of Y. lipolytica cells to the genomic stress in general and loss of telomerase in particular. Interestingly, Δpyl1 mutants did not exhibit increased sensitivity to DNA-damaging agents known to activate PARylation in mammalian cells, indicating that compared with mammalian PARPs Pyl1p plays a different role in the maintenance of genome stability. Based on our observations, we propose Pyl1 and its homologs we identified in other yeast species represent a distinct class of PARPs thus substantiating a more detailed investigation of their roles in these organisms.