biostudies-literatureTakeuchi KNCI NIH HHSNINDS NIH HHS886-899.e4https://www.ebi.ac.uk/biostudies/studies/S-EPMC9177683biostudies-literatureUnknown30(6)Unlike most kinases, phosphatidylinositol 5-phosphate 4-kinase β (PI5P4Kβ) utilizes GTP as a physiological phosphate donor and regulates cell growth under stress (i.e., GTP-dependent stress resilience). However, the genesis and evolution of its GTP responsiveness remain unknown. Here, we reveal that PI5P4Kβ has acquired GTP preference by generating a short dual-nucleotide-recognizing motif called the guanine efficient association (GEA) motif. Comparison of nucleobase recognition with 660 kinases and 128 G proteins has uncovered that most kinases and PI5P4Kβ use their main-chain atoms for adenine recognition, while the side-chain atoms are required for guanine recognition. Mutational analysis of the GEA motif revealed that the acquisition of GTP reactivity is accompanied by an extended actiStructure (London, England : 1993)The GTP responsiveness of PI5P4Kβ evolved from a compromised trade-off between activity and specificity.PMC9177683R01 NS089815R01 CA255331R21 NS100077Ikeda YOkuwaki KSasaki MTakeuchi KFukuzawa KImai MNagase LLo YHYu HYSasaki ATIto DOsaka NFujii YSenda THarada ANakagawa SSenda MfalseThe GTP responsiveness of PI5P4Kβ evolved from a compromised trade-off between activity and specificity.Unlike most kinases, phosphatidylinositol 5-phosphate 4-kinase β (PI5P4Kβ) utilizes GTP as a physiological phosphate donor and regulates cell growth under stress (i.e., GTP-dependent stress resilience). However, the genesis and evolution of its GTP responsiveness remain unknown. Here, we reveal that PI5P4Kβ has acquired GTP preference by generating a short dual-nucleotide-recognizing motif called the guanine efficient association (GEA) motif. Comparison of nucleobase recognition with 660 kinases and 128 G proteins has uncovered that most kinases and PI5P4Kβ use their main-chain atoms for adenine recognition, while the side-chain atoms are required for guanine recognition. Mutational analysis of the GEA motif revealed that the acquisition of GTP reactivity is accompanied by an extended acti2022-01-01T00:00:00Z2022 Jun2026-05-09T22:15:28.423Z2024-11-10T07:20:24.434ZS-EPMC91776833550427810.1016/j.str.2022.04.004