Poster abstracts
Poster number 61 submitted by Cydni Akesson
Investigating how DAZL regulates its RNA targets using BioID and immunostaining
Cydni Akesson (The Center for RNA Science & Therapeutics, Case Western Reserve University), Leah Zagore (The Center for RNA Science & Therapeutics, Case Western Reserve University), Mohammed Alzahrani (Case Western Reserve University), Molly Hannigan (The Center for RNA Science & Therapeutics, Case Western Reserve University), Donny Licatalosi (The Center for RNA Science & Therapeutics, Case Western Reserve University)
Abstract:
Deleted in Azospermia-Like (DAZL) is an RNA-binding protein expressed explicitly in germ cells. DAZL’s absence or malfunction causes infertility in male and female mice, and the molecular basis for this outcome is not well understood. Previous work in our lab indicates that DAZL positively regulates its RNA targets via 3’UTR interactions. To investigate how DAZL regulates its RNA targets, we will identify proteins interacting with DAZL using proximity-dependent biotin identification (BioID). We also use BioID and immunostaining to determine how different mutations in DAZL affect its ability to interact with different proteins and its subcellular localization. Thus far, we have found that DAZL’s protein network consists of many RNA processing factors and, although not exposed to stressful conditions in cell culture, many stress granule markers. These protein interactions are altered when point mutations are introduced into DAZL’s RNA Recognition Motif making it unable to bind RNA. Lastly, immunostaining experiments show a remarkable difference in DAZL’s localization in the cell depending on both its ability to bind RNA as well as the presence of its conserved, 27-amino acid intrinsically disordered region. Our results bring us closer to understanding how DAZL is able to access its target mRNAs in the cell.
Keywords: RNA Processing