NHR-23 and SPE-44 regulate distinct sets of genes during C. elegans spermatogenesis

ABSTRACT Spermatogenesis is the process through which mature male gametes are formed and is necessary for transmission of genetic information. While much work has established how sperm fate is promoted and maintained, less is known about how the sperm morphogenesis program is executed. We previously identified a novel role for the nuclear hormone receptor transcription factor, NHR-23, in promoting C. elegans spermatogenesis. Depletion of NHR-23 along with SPE-44, another transcription factor that promotes spermatogenesis, caused additive phenotypes. Through RNA-seq, we determined that NHR-23 and SPE-44 regulate distinct sets of genes. Depletion of both NHR-23 and SPE-44 produced yet another set of differentially regulated genes. NHR-23- regulated genes are enriched in phosphatases, consistent with the switch in spermatids to post-translational regulation following genome quiescence. In the parasitic nematode Ascaris suum , MFP1 and MFP2 control the polymerization of Major Sperm Protein, the molecule that drives sperm motility and serves as a signal to promote ovulation. NHR-23 and SPE-44 regulate a number of MFP2 paralogs, and NHR-23 depletion caused defective localization of MSD/MFP1 and NSPH-2/MFP2. Although NHR-23 and SPE-44 do not transcriptionally regulate the casein kinase gene spe-6, a key regulator of sperm development, SPE-6 protein is lost following NHR-23+SPE-44 depletion. Together, these experiments provide the first mechanistic insight into how NHR-23 promotes spermatogenesis and an entry point to understanding the synthetic genetic interaction between nhr-23 and spe-44 .