ArreSTick motif controls β-arrestin-binding stability and extends phosphorylation-dependent β-arrestin interactions to non-receptor proteins

• Published in: Cell reports (Cambridge) Vol. 43; no. 5; p. 114241
• Main Authors: Tóth, András Dávid, Soltész-Katona, Eszter, Kis, Katalin, Guti, Viktor, Gilzer, Sharon, Prokop, Susanne, Boros, Roxána, Misák, Ádám, Balla, András, Várnai, Péter, Turiák, Lilla, Ács, András, Drahos, László, Inoue, Asuka, Hunyady, László, Turu, Gábor
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 28.05.2024; Elsevier
• Subjects: Amino Acid Motifs; Amino Acid Sequence; arreSTick; arrestin; beta-Arrestin 2 - metabolism; beta-Arrestins - metabolism; convolution; CP: Molecular biology; GPCR; HEK293 Cells; HTATSF1; HTSF1; Humans; machine learning; mass spectrometry; Phosphorylation; Protein Binding; Protein Stability; proximity biotinylation assay; arreSTick; proximity biotinylation assay; CP: Molecular biology; convolution; GPCR; mass spectrometry; machine learning; arrestin; HTATSF1; HTSF1; phosphorylation
• ISSN: 2211-1247; 2211-1247
• DOI: 10.1016/j.celrep.2024.114241

The binding and function of β-arrestins are regulated by specific phosphorylation motifs present in G protein-coupled receptors (GPCRs). However, the exact arrangement of phosphorylated amino acids responsible for establishing a stable interaction remains unclear. We employ a 1D sequence convolution model trained on GPCRs with established β-arrestin-binding properties. With this approach, amino acid motifs characteristic of GPCRs that form stable interactions with β-arrestins can be identified, a pattern that we name “arreSTick.” Intriguingly, the arreSTick pattern is also present in numerous non-receptor proteins. Using proximity biotinylation assay and mass spectrometry analysis, we demonstrate that the arreSTick motif controls the interaction between many non-receptor proteins and β-arrestin2. The HIV-1 Tat-specific factor 1 (HTSF1 or HTATSF1), a nuclear transcription factor, contains the arreSTick pattern, and its subcellular localization is influenced by β-arrestin2. Our findings unveil a broader role for β-arrestins in phosphorylation-dependent interactions, extending beyond GPCRs to encompass non-receptor proteins as well. [Display omitted] •ArreSTick is a sequence pattern characteristic of stable GPCR-β-arrestin interaction•Numerous other proteins possess the arreSTick pattern•β-arrestin2 regulates HTSF1 via binding to its arreSTick motif Tóth et al. developed a machine learning-based approach to identify sequence motifs in GPCRs, termed arreSTick, that mediate stable β-arrestin binding. They found that arreSTick is also present in many other proteins, suggesting that β-arrestins may regulate non-GPCR proteins through a phosphorylation-dependent mechanism, similar to its function in GPCRs.