The topoisomerase II/condensin II axis silences transcription during germline specification in Caenorhabditis elegans

Abstract In Caenorhabditis elegans, the germline is specified via a preformation mechanism that relies on the PIE-1 protein's ability to globally silence mRNA transcription in germline precursor cells, also known as the P lineage. Recent work from our group has identified additional genome silencing events in C. elegans during oogenesis and in starved L1 larvae, and these require the condensin II complex, topoisomerase II, and components of the H3K9me/heterochromatin pathway. Interestingly, silencing in oocytes also requires PIE-1, but this is not the case in starved L1s. Here, we ask if additional genome silencing components besides PIE-1 are required to repress gene expression in the P lineage of early embryos, and we find that condensin II and topoisomerase II are required and the H3K9me/heterochromatin pathway is not. We show that depletion of topoisomerase II/condensin II activates the normally suppressed RNA polymerase II to inappropriately transcribe somatic genes in the P lineage. We also present evidence that while both PIE-1 and topoisomerase II/condensin II are required for genome silencing in the P lineage, PIE-1 can silence transcription independently of topoisomerase II/condensin II when misexpressed in somatic cells. Thus, in oocytes, all three genome silencing systems (topoisomerase II/condensin II, H3K9me, and PIE-1) are operational, while in both early embryos and starved L1s two of the three are active. Our data show that multiple, redundantly acting genome silencing mechanisms act in a mix-and-match manner to repress transcription at different developmental stages in the C. elegans germline.