title: The C terminus of the large tegument protein pUL36 contains multiple capsid binding sites that function differently during assembly and cell entry of herpes simplex virus.
abstract: The largest tegument protein of herpes simplex virus type 1 (HSV1), pUL36, is a multivalent cross-linker between the viral capsids and the tegument and associated membrane proteins during assembly that upon subsequent cell entry releases the incoming capsids from the outer tegument and viral envelope. Here we show that pUL36 was recruited to cytosolic progeny capsids that later colocalized with membrane proteins of herpes simplex virus type 1 (HSV1) and the trans-Golgi network. During cell entry, pUL36 dissociated from viral membrane proteins but remained associated with cytosolic capsids until arrival at the nucleus. HSV1 UL36 mutants lacking C-terminal portions of increasing size expressed truncated pUL36 but could not form plaques. Cytosolic capsids of mutants lacking the C-terminal 735 of the 3,164 amino acid residues accumulated in the cytosol but did not recruit pUL36 or associate with membranes. In contrast, pUL36 lacking only the 167 C-terminal residues bound to cytosolic capsids and subsequently colocalized with viral and host membrane proteins. Progeny virions fused with neighboring cells, but incoming capsids did not retain pUL36, nor could they target the nucleus or initiate HSV1 gene expression. Our data suggest that residues 2430 to 2893 of HSV1 pUL36, containing one binding site for the capsid protein pUL25, are sufficient to recruit pUL36 onto cytosolic capsids during assembly for secondary envelopment, whereas the 167 residues of the very C terminus with the second pUL25 binding site are crucial to maintain pUL36 on incoming capsids during cell entry. Capsids lacking pUL36 are targeted neither to membranes for virus assembly nor to nuclear pores for genome uncoating.
authors.affiliation: Institute of Virology, Hannover Medical School, Hannover, Germany.
authors.forename: Julia
authors.initials: J
authors.lastname: Schipke
authors.forename: Anja
authors.initials: A
authors.lastname: Pohlmann
authors.forename: Randi
authors.initials: R
authors.lastname: Diestel
authors.forename: Anne
authors.initials: A
authors.lastname: Binz
authors.forename: Kathrin
authors.initials: K
authors.lastname: Rudolph
authors.forename: Claus-Henning
authors.initials: CH
authors.lastname: Nagel
authors.forename: Rudolf
authors.initials: R
authors.lastname: Bauerfeind
authors.forename: Beate
authors.initials: B
authors.lastname: Sodeik
mesh_terms: D020816:Amino Acid Motifs; D000818:Animals; D001665:Binding Sites; D036022:Capsid Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D006561:Herpes Simplex / Q000821:virology*; D018259:Herpesvirus 1, Human / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D006801:Humans; D011485:Protein Binding; D014764:Viral Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D019065:Virus Assembly; D053586:Virus Internalization; D021601:trans-Golgi Network / Q000821:virology
chemical_list: D036022:Capsid Proteins; C078035:UL36 protein, Human herpesvirus 1; D014764:Viral Proteins