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Deletion or green fluorescent protein tagging of the pUL35 capsid component of pseudorabies virus impairs virus replication in cell culture and neuroinvasion in mice

Institute of Molecular Biology, Friedrich-Loeffler-Institut, 17493 Greifswald-Insel Riems, Germany

Correspondence
Walter Fuchs
walter.fuchs{at}fli.bund.de

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.

Supplementary material is available with the online version of this paper.

This article has been cited by other articles:

				M. Krautwald, W. Fuchs, B. G. Klupp, and T. C. Mettenleiter Translocation of Incoming Pseudorabies Virus Capsids to the Cell Nucleus Is Delayed in the Absence of Tegument Protein pUL37 J. Virol., April 1, 2009; 83(7): 3389 - 3396. [Abstract] [Full Text] [PDF]