Puf4 is methylated and exhibits a temperature-dependent interactome in

The evolutionarily conserved Pumilio domain-containing RNA binding proteins (RBPs) are involved in many steps of post-transcriptional gene regulation, including RNA stability, polyadenylation, deadenylation, and translation. Most RBPs are post-translationally modified by methylation of arginine/glycine-rich domains (RG-box) though the consequences of these modifications are not well known. We determined the arginine methylation and phosphorylation landscape of the Pumilio domain containing RBP Puf4 from the basidiomycete fungus IMPORTANCE: Post-transcriptional processes play a crucial role in modulating responses to various stressors, including temperature and antifungals. To ensure adaptation and survival, stressors must be accurately sensed, and responses tightly regulated. Post-translational modifications serve as a crucial regulatory layer that orchestrates nuanced stress responses. We highlight extensive methylation and phosphorylation of the RNA-binding protein Puf4 and explore the regulatory roles of select methylated arginines. We also found that the interactome network of Puf4 varies with temperature. Cryptococcus neoformans, a basidiomycete fungus that is also an important human pathogen, is an ideal system to illustrate the evolution of gene regulatory processes and how these kingdom or clade-specific regulatory modules contribute to the pathogenic potential of fungi.