Makorin 1 controls embryonic patterning by alleviating Bruno1-mediated repression ofoskartranslation

Abstract

AbstractMakorins are evolutionary conserved proteins that contain C3H-type zinc finger modules and a RING E3 ubiquitin ligase domain. InDrosophilamaternal Makorin 1 (Mkrn1) has been linked to embryonic patterning but the mechanism remained unsolved. Here, we show that Mkrn1 is essential for axis specification and pole plasm assembly by translational activation ofoskar. We demonstrate that Mkrn1 interacts with poly(A) binding protein (pAbp) and bindsosk3’ UTR in a region adjacent to A-rich sequences. This binding site overlaps with Bruno1 (Bru1) responsive elements (BREs), which regulateosktranslation. We observe increased association of the translational repressor Bru1 withoskmRNA upon depletion of Mkrn1, indicating that both proteins compete foroskbinding. Consistently, reducing Bru1 dosage partially rescues viability and Osk protein level in ovaries fromMkrn1females. We conclude that Mkrn1 controls embryonic patterning and germ cell formation by specifically activatingosktranslation by displacing Bru1 from its 3’ UTR.Author SummaryTo ensure accurate development of theDrosophilaembryo, proteins and mRNAs are positioned at specific sites within the embryo. Many of these proteins and mRNAs are produced and localized during the development of the egg in the mother. One protein essential for this process that has been heavily studied is Oskar (Osk), which is positioned at the posterior pole. During the localization ofoskmRNA, its translation is repressed by the RNA-binding protein Bruno1 (Bru1), ensuring that Osk protein is not present outside of the posterior where it is harmful. At the posterior pole,oskmRNA is activated through mechanisms that are not yet understood. In this work, we show that the conserved protein Makorin 1 (Mkrn1) is a novel factor involved in the translational activation ofosk. Mkrn1 binds specifically tooskmRNA in a region that overlaps with the binding site of Bru1, thus alleviating the association of Bru1 withosk. Moreover, Mkrn1 is stabilized by poly(A) binding protein, a translational activator that bindsoskmRNA in close proximity to Mkrn1. Our work thus helps to answer a long-standing question in the field, providing insight about the function of Mkrn1 and more generally into embryonic patterning in animals.