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

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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, 133 (21). jcs249664. ISSN 0021-9533 (https://doi.org/10.1242/jcs.249664)

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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 cysteine residues 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 cysteine-265 and -268 were identified as key targets of this enzyme. In contrast, the low specificity/high activity zDHHC3 and 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 α-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.

ORCID iDs

Locatelli, Carolina, Lemonidis, Kimon ORCID logoORCID: https://orcid.org/0000-0003-1332-6002, Salaun, Christine ORCID logoORCID: https://orcid.org/0000-0003-3002-6810, Tomkinson, Nicholas C. O. ORCID logoORCID: https://orcid.org/0000-0002-5509-0133 and Chamberlain, Luke H. ORCID logoORCID: https://orcid.org/0000-0002-8701-4995;
CORE (COnnecting REpositories)