Altering a gene involved in nuclear distribution increases the repeat-induced point mutation process in the fungus Podospora anserina

Repeat-induced point mutation (RIP) is a homology-dependent gene-silencing mechanism that introduces C:G-to-T:A transitions in duplicated DNA segments. Cis-duplicated sequences can also be affected by another mechanism called premeiotic recombination (PR). Both are active over the sexual cycle of some filamentous fungi, e.g., Neurospora crassa and Podospora anserina. During the sexual cycle, several developmental steps require precise nuclear movement and positioning, but connections between RIP, PR, and nuclear distributions have not yet been established. Previous work has led to the isolation of amil, the P. anserina ortholog of the Aspergillus nidulans apsA gene, which is required for nuclear positioning. We show here that amil is involved in nuclear distribution during the sexual cycle and that alteration of amil delays the fruiting-body development. We also demonstrate that amil alteration affects loss of transgene functions during the sexual cycle. Genetically linked multiple copies of transgenes are affected by RIP and PR much more frequently in an amil mutant cross than in a wild-type cross. Our results suggest that the developmental slowdown of the amil mutant during the period of RIP and PR increases time exposure to the duplication detection system and thus increases the frequency of RIP and PR.