Protein synthesis and degradation are essential to regulate germline stem cell homeostasis in Drosophila testes

The homeostasis of self-renewal and differentiation in stem cells is strictly controlled by intrinsic signals and their niche. We conducted a large-scale RNA interference (RNAi) screen in <i>Drosophila</i> testes and identified 221 genes required for germline stem cell (GSC) maintenance or differentiation. Knockdown of these genes in transit-amplifying spermatogonia and cyst cells further revealed various phenotypes. Complex analysis uncovered that many of the identified genes are involved in key steps of protein synthesis and degradation. A group of genes that are required for mRNA splicing and protein translation contributes to both GSC self-renewal and early germ cell differentiation. Loss of genes in protein degradation pathway in cyst cells leads to testis tumor with overproliferated germ cells. Importantly, in the Cullin 4-Ring E3 ubiquitin ligase (CRL4) complex, we identified multiple proteins that are critical to GSC self-renewal. pic/DDB1, the linker protein of CRL4, is not only required for GSC self-renewal in flies but also for maintenance of spermatogonial stem cells (SSCs) in mice.