Investigation of herpes simplex virus type 1 genes encoding multiply inserted membrane proteins

Abstract

The herpes simplex virus type 1 genome contains four open reading frames (ORFs) which are predicted to encode hydrophobic proteins with the potential to cross a membrane several times. The products of these genes (genes UL10, UL20, UL43 and UL53) have not previously been identified. To investigate the role of these proteins in the virus life cycle, we attempted to inactivate the genes individually by inserting the lacZ gene from Escherichia coli within the ORFs. Using this approach we have isolated insertion mutants for UL10 and UL43, as well as a deletion mutant lacking the majority of the UL43 ORF. The growth of the UL10-lacZ virus was slightly impaired in tissue culture compared to that of the wild-type virus parent, whereas the growth of the UL43 mutants was indistinguishable from that of wild-type virus. Furthermore, deletion of the majority of the UL43 ORF did not impair the ability of the virus to replicate in vivo at the periphery, or to spread to and replicate within the nervous system, in a mouse ear model. Repeated attempts to isolate lacZ insertion mutants for UL20 and UL53 were unsuccessful, suggesting that these genes may be essential for virus growth, at least in tissue culture. Using antipeptide sera, the products of genes UL10 and UL20 have been detected.

Time-resolved Global and Chromatin Proteomics during Herpes Simplex Virus Type 1 (HSV-1) Infection

“…Two of the late genes, UL43 and US8.5, were detected only at 6 hpi and 12hpi. The function of both proteins is still unknown, although UL43 encodes a hydrophobic, myristoylated integral membrane protein (76,77), whereas US8.5 encodes a nuclear protein that localizes to nucleoli and its mRNA is among the most abundant species packaged in virions (78). The protein intensities of US8.5 and UL43 were ϳ40 and 270-fold (respectively) lower than the average intensity of all detected HSV-1 proteins.…”

Selective astrocytic transgene expression in vitro and in vivo from the GFAP promoter in a HSV RL1 null mutant vector – potential glioblastoma targeting

“…The parental wild-type HSV-1 strain used in this study was Glasgow strain 17 + , 30 and the parental RL1 mutant was 1716. 10 1716pR9 23 and 1716SV40 31 (AR MacLean, personal communication) were used as positive controls for some experiments. These viruses contain the LAT P1 and SV40 promoters, respectively, linked to a ␤-galactosidase reporter gene and inserted in the HSV-1 UL43 locus.…”

“…31 This plasmid has been modified by insertion of a double stranded oligonucleotide containing a multicloning site with a unique BglII site at the unique NsiI site (np 94911) within UL43 and was provided by Dr A McGregor (Division of Virology, Glasgow University, UK). A 5.2 kb BamHI fragment from the plasmid pGFa2l-acZ, 18 containing a 2.2 kb fragment from the GFAP promoter linked to a ␤-galactosidase reporter gene was cloned into the unique BglII site in p35 to produce the plasmid p35-GFAP.…”

Deletion of glycoprotein gM of pseudorabies virus results in attenuation for the natural host.

“…gB, gH and gL are necessary for productive replication in all viruses analysed in this respect. In contrast, gM is dispensable for replication of herpes simplex virus 1 (HSV-1 ; Baines & Roizman, 1991MacLean et al, 1991MacLean et al, , 1993, equine herpesvirus 1 (Osterrieder et al, 1996) and PrV (Dijkstra et al, 1996). In these viruses, lack of gM resulted in a decrease of replicative ability in cell culture represented by 10-to 50-fold lower virus titres, a delay in entry into target cells, and slightly reduced plaque size.…”