A lysine-methionine exchange in a coronavirus surface protein transforms a retention motif into an endocytosis signal.

Transmissible gastroenteritis virus (TGEV) is an enveloped (+) RNA virus belonging to the family Coronaviridae. Among the viral membrane proteins, the spike (S) protein mediates receptor recognition/attachment to the host cell and fusion of viral and cellular membranes. The cytoplasmic tail of the S protein contains a tyrosine-dependent sorting signal with the consensus sequence YXXΦ. In the context of the S protein of TGEV (1440YEPI1443), this motif acts as a retention signal, preventing surface expression of the protein. Here, we show that a chimeric S protein, containing the six C-terminal amino acids of the glycoprotein G of vesicular stomatitis virus (VSV) is no longer retained intracellularly, despite the presence of the tyrosine tetrapeptide motif. Following transport to the cell surface, the chimeric protein was rapidly endocytosed. Analysis of mutant proteins generated by site-directed mutagenesis revealed that a single amino acid exchange (1445K/M, position: +2 downstream of the tyrosine-based motif) was responsible for the altered sorting behavior.