Striebinger, HannahHannahStriebingerZhang, JieJieZhangOtt, MelanieMelanieOttFunk, ChristinaChristinaFunkRadtke, KerstinKerstinRadtkeDuron, JohanneJohanneDuronRuzsics, ZsoltZsoltRuzsicsHaas, JürgenJürgenHaasLippé, RogerRogerLippéBailer, SusanneSusanneBailer2022-03-052022-03-052015https://publica.fraunhofer.de/handle/publica/24296510.1099/jgv.0.000262The Herpes simplex virus type 1 (HSV-1) glycoprotein M (gM/UL10) is a 473 amino acid type III transmembrane protein that resides in various membrane compartments. HSV-1 gM contains several putative trafficking motifs but their functional relevance remains to be elucidated. We show here that transiently expressed gM 19-343 was sufficient for transport to the TGN, while gM 133-473 where the first two transmembrane domains were deleted, and gM 1-342 that lacked the final residue of the last transmembrane domain, were retained in the ER indicating that all transmembrane domains are required for proper folding and ER exit. A series of BAC mutants revealed that in addition to the authentic start codon, translation of gM can be initiated at methionine 19 and 133/135. While a protein lacking the first 18 residues supported wild type-like growth, gM 133/135-473 resulted in reduced plaque diameters resembling a UL10 deletion mutant. An HSV-1 mutant encoding gM 1-342 showed similar growth characteristics and accumulated un-enveloped cytoplasmic particles while gM 1-343 resulted in a gain of function, indicating that all transmembrane domains of the protein are important for viral growth. A C-terminal extension further supported viral propagation, the C-terminal trafficking motifs (residues 423-473) however were completely dispensable. We propose a functional core within gM 19-343 comprised of all transmembrane domains that is sufficient to target the protein to the TGN, a favoured site for envelopment, and to support viral functions.en579journal article