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DiNardo, A. L.

Publications and source records attributed to DiNardo, A. L..

2 recordsLinked to original sources

Caenorhabditis elegans form sexually dimorphic and dynamic germ granules throughout meiotic prophase I progression

In sexually reproducing organisms, germ cells faithfully transmit both the genome and epigenetic information across generations through the formation of haploid gametes, such as eggs and sperm. Small RNA pathways tune gene expression in a sex-specific manner during germ cell development to facilitate both proper germ cell formation and transgenerational inheritance of epigenetic information. In Caenorhabditis elegans, components of small RNA pathways localize to germ granules, liquid-like membraneless organelles within the cytoplasm of developing germ cells. During oogenesis, germ granules form hierarchal sub-compartments that may be required for proper germ cell development and epigenetic inheritance. However, germ granule structure during spermatogenesis remains largely undescribed. Here we determine that the germ granule structural components PGL-1 and ZNFX-1 display sexually dimorphic foci morphology and size during meiotic prophase I progression. Further, we quantitate the sexually dimorphic sub-compartmentalization of these two proteins within the germ granule, determining that while PGL-1 and ZNFX-1 do associate during germ cell development, the extent of overlap varies between sexes and throughout meiotic progression. Additionally, we identify WAGO-4, a Argonaute protein central to gene regulation by small RNA pathways, as a sexually dimorphic component of the germ granule during germ cell development. Together, our studies reveal that the overall structure of the germ granule, as well as an Argonaute protein housed inside, are sexually dimorphic, which may underpin sex-specific regulation by small RNA pathways during germ cell development. Author SummarySmall RNA pathways are critical for the regulation and passage of genomic and epigenetic information to the next generation. Components of these pathways are housed in germ granules during egg and sperm development. Previous work examining germ granule structure in Caenorhabditis elegans focused on oocytes and the late stages of meiosis I. Here, we comprehensively characterize the localization of two structural and one functional component of the germ granule throughout meiotic progression of both developing egg and sperm cells. We identify that both biophysical properties and germ granule configurations are dependent upon meiotic stage and sex.

developmental biology↗

WAGO-1 is a sexually dimorphic Argonaute protein required for proper germ granule structure and gametogenesis

Germ cell proliferation and proper genome inheritance are critical for maintaining fertility through generations. To promote proper germ cell development, small RNA pathways employ Argonaute proteins (AGOs) to modulate gene expression and protect against deleterious genomic elements while not silencing against self. Here we identify sexual dimorphisms in localization and function of protein structural features of the Argonaute WAGO-1 that affects sex-specific gene regulation during C. elegans germ cell development. During meiotic prophase I progression, we find that germ granule structural proteins and the PIWI AGO, PRG-1, display dynamic and distinct localization patterns between egg and sperm development which coincide with differential WAGO-1 localization and biophysical properties. Sexually dimorphic functions of specific WAGO-1 protein structural domains underpin these differences. Disruption or modification to the N-terminus intrinsically disordered region (IDR) of WAGO-1 leads to loss of PGL-1 phase separation only during spermatogenesis. Further, we find that these germ granule disruptions are likely due to prolonged association of the IDR with the RNA-binding pocket of WAGO-1. In addition, deletion of the MID and part of the PIWI domains causes male-specific sterility and disruption to WAGO-1 localization with PGL-1 during oogenesis. Finally, we demonstrate that these disruptions to WAGO-1 protein structure dynamically change the mRNA and sRNA landscape of adult males and hermaphrodites, in which the AGOs ALG-3/4 and VSRA-1 are misregulated. Together, these data suggest that WAGO-1 differentially regulates genes during oogenesis versus spermatogenesis, and that these differences in gene regulation may be due to the sex-specific configuration and biophysical properties of WAGO-1 within the germ granule.

developmental biology↗