segmental duplications arise from pol32-dependent repair of broken forks through two alternative replication-based mechanisms节段复制来自pol32-dependent修复破碎的叉通过两种replication-based机制.pdfVIP

Segmental Duplications Arise from Pol32-Dependent Repair of Broken Forks through Two Alternative Replication-Based Mechanisms Celia Payen., Romain Koszul.¤, Bernard Dujon, Gilles Fischer* ´ ´ ´ ´ ´ Institut Pasteur, Unite de Genetique Moleculaire des Levures, CNRS, URA2171, Universite Pierre et Marie Curie-Paris 6, UFR927, Paris, France Abstract The propensity of segmental duplications (SDs) to promote genomic instability is of increasing interest since their involvement in numerous human genomic diseases and cancers was revealed. However, the mechanism(s) responsible for their appearance remain mostly speculative. Here, we show that in budding yeast, replication accidents, which are most likely transformed into broken forks, play a causal role in the formation of SDs. The Pol32 subunit of the major replicative polymerase Pold is required for all SD formation, demonstrating that SDs result from untimely DNA synthesis rather than from unequal crossing-over. Although Pol32 is known to be required for classical (Rad52-dependant) break-induced replication, only half of the SDs can be attributed to this mechanism. The remaining SDs are generated through a Rad52- independent mechanism of template switching between microsatellites or microhomologous sequences. This new mechanism, named microhomology/microsatellite-induced replication (MMIR), differs from all known DNA double-strand break repair pathways, as MMIR-mediated duplications still occur in the combined absence of homologous recombination, microhomology-mediated, and nonhomologous end joining machineries. The interplay between these two replication- based pathways explains important features of higher eukaryotic genomes,