A conserved motif in Pch2 regulates its localization and meiotic function in Saccharomyces cerevisiae

The Saccharomyces cerevisiae Pch2 protein is a conserved meiotic AAA+ ATPase whose activity must be tightly regulated to ensure proper chromosome dynamics during meiotic prophase I. Its function relies on remodeling the HORMA-domain protein Hop1, promoting conformational transitions that are essential for chromosome axis organization, checkpoint signaling, and recombination control. Here, we identify threonine 428 (T428), located within a conserved threonine-glutamine (TQ) putative phosphorylation motif, as a critical regulatory residue of Pch2. We found that, in zip1Δ cells, the meiotic recombination checkpoint response is partially or completely abolished in the pch2-T428A and pch2-T428D mutants, respectively. Both mutations alter Pch2 subcellular localization, leading to its increased nuclear accumulation; however, forced nuclear exclusion of Pch2-T428A, but not Pch2-T428D, restores the zip1Δ meiotic block, indicating an additional effect of the T428D substitution on checkpoint function beyond subcellular distribution. Analysis in synapsis-proficient strains reveals that this residue also plays a critical role in coordinating Hop1 chromosomal enrichment with Mek1 activation along the synaptonemal complex. In contrast to pch2Δ or the ATPase-defective pch2-E399Q mutant, introduction of a negative charge at the 428 position uncouples Hop1 accumulation from its phosphorylation, preventing Mek1 activation despite robust Hop1 association with meiotic chromosomes. These findings support emerging models in which Pch2 regulates Hop1 to control not only its chromosomal abundance, but also the maintenance of sufficient levels of Hop1 in a phosphorylation-competent conformation, thereby ensuring proper checkpoint signaling and faithful meiotic progression.