Identification of key features required for efficient S-acylation and plasma membrane targeting of Sprouty-2

Locatelli, Carolina and Lemonidis, Kimon and Salaun, Christine and Tomkinson, Nicholas C.O. and Chamberlain, Luke H. (2020) Identification of key features required for efficient S-acylation and plasma membrane targeting of Sprouty-2. Journal of Cell Science. ISSN 0021-9533 (In Press)

[img] Text (Locatelli-etal-JCS-2020-Identification-of-key-features-required-for-efficient-S-acylation)
Locatelli_etal_JCS_2020_Identification_of_key_features_required_for_efficient_S_acylation.pdf
Accepted Author Manuscript
Restricted to Repository staff only until 30 September 2021.

Download (10MB) | Request a copy from the Strathclyde author

    Abstract

    Sprouty-2 is an important regulator of growth factor signalling and a tumour suppressor protein. The defining feature of this protein is a cysteine-rich domain (CRD) that contains twenty-six cysteines and is modified by S-acylation. In this study, we show that the CRD of Sprouty-2 is differentially modified by S-acyltransferase enzymes. The high specificity/low activity zDHHC17 enzyme mediated restricted S-acylation of Sprouty-2, and cysteines-265/268 were identified as key targets of this enzyme. In contrast, the low specificity/high activity zDHHC3/zDHHC7 enzymes mediated more expansive modification of the Sprouty-2 CRD. Nevertheless, S-acylation by all enzymes enhanced Sprouty-2 expression, suggesting that S-acylation stabilises this protein. In addition, we identified two charged residues (aspartate-214 and lysine-223), present on opposite faces of a predicted alpha helix in the CRD, which are essential for S-acylation of Sprouty-2. Interestingly, mutations that perturbed S-acylation also led to a loss of plasma membrane localisation of Sprouty-2 in PC12 cells. This study provides insight into the mechanisms and outcomes of Sprouty-2 S-acylation, and highlights distinct patterns of S-acylation mediated by different classes of zDHHC enzymes.