The endoplasmic reticulum-localized enzyme zDHHC6 mediates S-acylation of short transmembrane constructs from multiple type I and II membrane proteins

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Salaun, Christine and Tomkinson, Nicholas C.O. and Chamberlain, Luke H. (2023) The endoplasmic reticulum-localized enzyme zDHHC6 mediates S-acylation of short transmembrane constructs from multiple type I and II membrane proteins. Journal of Biological Chemistry, 299 (10). 105201. ISSN 1083-351X (https://doi.org/10.1016/j.jbc.2023.105201)

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Abstract

In this study, we investigated the S-acylation of two host cell proteins important for viral infection: TMPRSS2 (transmembrane serine protease 2), which cleaves SARS-CoV-2 spike to facilitate viral entry, and BST2 (Bone marrow stromal antigen 2), a general viral restriction factor. We found that both proteins were S-acylated by zDHHC6, an S-acyltransferase enzyme localized at the endoplasmic reticulum (ER), in co-expression experiments. Mutagenic analysis revealed that zDHHC6 modifies a single cysteine in each protein, which are in proximity to the transmembrane domains (TMDs). For TMPRSS2, the modified cysteine is positioned two residues into the TMD, whereas the modified cysteine in BST2 has a cytosolic location two amino acids upstream of the TMD. Cysteine swapping revealed that repositioning the target cysteine of TMPRSS2 further into the TMD substantially reduced S-acylation by zDHHC6. Interestingly, zDHHC6 efficiently S-acylated truncated forms of these proteins that contained only the TMDs and short juxtamembrane regions. The ability of zDHHC6 to modify short TMD sequences was also seen for the transferrin receptor (another type II membrane protein) and for five different type I membrane protein constructs, including CD4. Collectively, the results of this study show that zDHHC6 can modify diverse membrane proteins (Type I and II) and requires only the presence of the TMD and target cysteine for efficient S-acylation. Thus, zDHHC6 may be a broad specificity S-acyltransferase specialized for the modification of a diverse set of transmembrane proteins at the endoplasmic reticulum.

ORCID iDs

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)