Zebrafish reveal new roles for Fam83f in hatching and the DNA damage-mediated autophagic response

Abstract

AbstractThe FAM83 (Family with sequence similarity83) family is highly conserved in vertebrates, but little is known of the functions of these proteins beyond their association with oncogenesis. Of the family, FAM83F is of particular interest because it is the only membrane-targeted FAM83 protein. When over-expressed, FAM83F activates the canonical Wnt signalling pathway and binds to and stabilizes p53; it therefore interacts with two pathways often dysregulated in disease. Insights into gene function can often be gained by studying the roles they play during development, and here we report the generation offam83fknock-out (KO) zebrafish, which we have used to study the role of Fam83f in vivo. We show that endogenousfam83fis most strongly expressed in the hatching gland of developing zebrafish embryos, and thatfam83fKO embryos hatch earlier than their wild-type (WT) counterparts, despite developing at a comparable rate. We also demonstrate thatfam83fKO embryos are more sensitive to ionizing radiation than WT embryos—an unexpected finding, bearing in mind the previously-reported ability of FAM83F to stabilize p53. Transcriptomic analysis shows that loss offam83fleads to downregulation of phosphatidylinositol-3-phosphate (PI(3)P) binding proteins and impairment of cellular degradation pathways, particularly autophagy, a crucial component of the DNA damage response. Finally, we show that Fam83f protein is itself targeted to the lysosome when over-expressed in HEK293T cells, and that this localization is dependent upon a C’ terminal signal sequence. The zebrafish lines we have generated suggest that Fam83f plays an important role in autophagic/lysosomal processes, resulting in dysregulated hatching and increased sensitivity to genotoxic stress in vivo.