Transglutaminase 2 (TG2) is a diverse, ubiquitous, multifunctional protein involved in diverse biological functions. Evidence from our previous study depicts TG2 binds RNA, potentially playing a role in post-transcriptional regulations. The objective of this study was to decipher the RNA-binding site within transglutaminase 2 and to demonstrate the amino acid residues that are involved in this interaction. Our results demonstrated that truncated TG2 (1-582aa) maintained its RNA binding ability, suggesting that the C-terminal beta-barrel domain is not involved in the RNA binding. However, TG2 nucleotide binding site mutants demonstrated impaired RNA-binding properties suggesting that the GTP binding sites present on the core domain within TG2 contributes to TG2- RNA interaction. In conclusion, our results provide additional evidence about the TG2 RNA binding site and its potential regulation by nucleotide binding, which could be involved in the mechanism of essential cellular processes.