Abstract
The germ plasm has long been demonstrated to be necessary and sufficient for germline determination, with translational regulation playing a key role in the process. Beyond this, little is known about molecular activities underlying germline determination.We report the function of Drosophila PIWI, DICER-1, and dFMRP (Fragile X Mental Retardation Protein) in germline determination. PIWI is a maternal component of the polar granule, a germ-plasm-specific organelle essential for germline specification. Depleting maternal PIWI does not affect OSK or VASA expression or abdominal patterning but leads to failure in pole-plasm maintenance and primordial-germ-cell (PGC) formation, whereas doubling and tripling the maternal piwi dose increases OSK and VASA levels correspondingly and doubles and triples the number of PGCs, respectively. Moreover, PIWI forms a complex with dFMRP and DICER-1, but not with DICER-2, in polar-granule-enriched fractions. Depleting DICER-1, but not DICER-2, also leads to a severe pole-plasm defect and a reduced PGC number. These effects are also seen, albeit to a lesser extent, for dFMRP, another component of the miRISC complex.Because DICER-1 is required for the miRNA pathway and DICER-2 is required for the siRNA pathway yet neither is required for the rasiRNA pathway, our data implicate a crucial role of the PIWI-mediated miRNA pathway in regulating the levels of OSK, VASA, and possibly other genes involved in germline determination in Drosophila.
