A distinct mechanism of RNA recognition by the transcription factor GATA1

ABSTRACT Several human transcription factors (TFs) have been reported to directly bind RNA through non-canonical RNA-binding domains; however, most of these TFs remain to be further validated as bona fide RNA-binding proteins (RBPs). Our systematic analysis of RBP discovery datasets reveals a varied set of candidate TF-RBPs that encompass most TF families. These candidate RBPs include members of the GATA family, which are essential factors in embryonic development. Investigation of the RNA-binding features of GATA1, a major hematopoietic TF, reveals robust non-sequence specific binding to RNAs in vitro . Moreover, RNA binding by GATA1 is competitive with DNA binding, which occurs through a shared binding surface spanning the DNA-binding domain and arginine-rich motif (ARM) like domain. We show that the ARM-like domain contributes both substantially to high-affinity DNA binding and electrostatically to plastic RNA recognition, suggesting that the separable RBD assigned to the ARM-domain in GATA1 is an oversimplification of a more complex recognition network. These biochemical data demonstrate a unified integration of DNA- and RNA-binding surfaces within GATA1, whereby the ARM-like domain provides an electrostatic surface for RNA binding but does not fully dominate GATA1-RNA interactions, which may also apply to other TF-RBPs. This competitive DNA/RNA binding activity using overlapping nucleic acid binding regions points to the possibility of RNA-mediated regulation of GATA1 function during hematopoiesis. Our study highlights the multifunctionality of DNA-binding domains in RNA recognition and supports the need for robust characterization of predicted non-canonical RNA-binding domains such as ARM-like domains.