Post-transcriptional gene regulation by RNA-binding proteins has proved crucial for developmental events including establishment and patterning of body axes, asymmetric cell fate decisions, and morphogenesis. Among RNA-binding proteins, heterogeneous nuclear ribonucleoproteins (hnRNPs) are notable for their ability to interact with numerous target RNAs and function in multiple aspects of RNA metabolism. The basis for this functional diversity is poorly understood, however. We previously identified Glo as an ovarian repressor of nanos (nos) translation through its interaction with a double-stranded UA-rich motif in the nanos 3' untranslated region (3'UTR). In contrast, human hnRNP F/H proteins are best known for their role in the regulation of alternative splicing through their interaction with single-stranded G-tract sequences. Consistent with this, we have also uncovered a potential role for Glo as a splicing factor. To investigate the multifunctionality of Glo, we determined the crystal structures of Glo’s three quasi-RNA recognition motifs (qRRMs) and performed a structure/function analysis. We found that Glo qRRMs recognize G-tracts similarly to hnRNP F but recognize the UA-rich motif in nos through a second, noncanonical RNA-binding interface. By assaying the effect of mutations that disrupt each recognition mode in vitro on Glo function in vivo, we demonstrated that regulation of a subset of Glo’s targets in vivo is mediated solely using the G-tract binding mode whereas regulation of nos requires both modes of recognition. This latter result led to the identification a G-tract in the nos 3'UTR that mediates Glo binding and is indeed required for nanos regulation. Finally, we establish that Glo’s requirement for both RNA binding modes to regulate nos translation reflects a requirement for both motifs in the nos 3'UTR.