title: Deletion or green fluorescent protein tagging of the pUL35 capsid component of pseudorabies virus impairs virus replication in cell culture and neuroinvasion in mice.
abstract: To facilitate tracing of virion movement, the non-essential capsid proteins pUL35 of herpes simplex virus type 1 and pseudorabies virus (PrV) have been tagged with green fluorescent protein (GFP). However, the biological relevance of PrV pUL35 and the functionality of the fusion proteins have not yet been investigated in detail. We generated PrV mutants either lacking the 12 kDa UL35 gene product, or expressing GFP fused to the N terminus of pUL35. Remarkably, both mutants exhibited significant replication defects in rabbit kidney cells, which could be corrected in pUL35-expressing cells. After intranasal infection of mice both mutants showed delayed neuroinvasion, and survival times of the animals were extended to 3 days, compared with 2 days after wild-type infection. Thus, fusion of pUL35 with GFP resulted in a non-functional protein, which has to be considered for the use of corresponding mutants in tracing studies.
authors.affiliation: Institute of Molecular Biology, Friedrich-Loeffler-Institut, 17493 Greifswald-Insel Riems, Germany.
authors.forename: Mirjam
authors.initials: M
authors.lastname: Krautwald
authors.affiliation: Institute of Molecular Biology, Friedrich-Loeffler-Institut, 17493 Greifswald-Insel Riems, Germany.
authors.forename: Christina
authors.initials: C
authors.lastname: Maresch
authors.affiliation: Institute of Molecular Biology, Friedrich-Loeffler-Institut, 17493 Greifswald-Insel Riems, Germany.
authors.forename: Barbara G
authors.initials: BG
authors.lastname: Klupp
authors.affiliation: Institute of Molecular Biology, Friedrich-Loeffler-Institut, 17493 Greifswald-Insel Riems, Germany.
authors.forename: Walter
authors.initials: W
authors.lastname: Fuchs
authors.affiliation: Institute of Molecular Biology, Friedrich-Loeffler-Institut, 17493 Greifswald-Insel Riems, Germany.
authors.forename: Thomas C
authors.initials: TC
authors.lastname: Mettenleiter
mesh_terms: D000818:Animals; D036022:Capsid Proteins / Q000502:physiology*; D002460:Cell Line; D004417:Dyspnea / Q000473:pathology; D049452:Green Fluorescent Proteins / Q000378:metabolism; D011558:Herpesvirus 1, Suid / Q000502:physiology*; D051379:Mice; D017354:Point Mutation; D011557:Pseudorabies / Q000473:pathology / Q000821:virology*; D011817:Rabbits; D014779:Virus Replication
chemical_list: D036022:Capsid Proteins; D049452:Green Fluorescent Proteins