Genome sequence of a human tumorigenic poxvirus: prediction of specific host response-evasion genes.

Molluscum contagiosum virus (MCV) commonly causes asymptomatic cutaneous neoplasms in children and sexually active adults as well as persistent opportunistic acquired immunodeficiency syndrome (AIDS)-associated disease. Sequencing the 190-kilobase pair genome of MCV has now revealed that the virus potentially encodes 163 proteins, of which 103 have homologs in the smallpox virus. MCV lacks counterparts to 83 genes of the smallpox virus, including those important in suppression of host responses to infection, nucleotide biosynthesis, and cell proliferation. MCV possesses 59 genes that are predicted to encode previously uncharacterized proteins, including major histocompatibility complex class I, chemokine, and glutathione peroxidase homologs, which suggests that there are MCV-specific strategies for coexistence with the human host.

Nucleotide sequence analysis of a cloned DNA fragment from human cells reveals homology to retrotransposons.

During molecular cloning of proviral DNA of human spumaretrovirus, various recombinant clones were established and analyzed. Blot hybridization revealed that one of the recombinant plasmids had the characteristic features of a member of the long interspersed repetitive sequences family. The DNA element was analyzed by restriction mapping and nucleotide sequencing. It showed a high degree of amino acid sequence homology of 54.3% when compared with the 5'-terminal part of the pol gene product of the murine retrotransposon LIMd. The 3' region of the cloned DNA element encodes proteins with an even higher degree of homology of 67.4% in comparison to the corresponding parts of a member of the primate KpnI sequence family.

The nucleotide sequence of the inverted terminal repetition of the tree shrew adenovirus DNA.

The termini of the tupaia (tree shrew) adenovirus (TAV) DNA have been sequenced. The inverted terminal repetitions (ITR) are 166 bp long containing the A + T-rich, highly conserved sequence present in all adenovirus DNAs so far analysed. An unusual feature within the TAV ITR is the presence of four sets of a conserved sequence TGACCG which occur at or near the ends of many adenovirus ITR.

Molecular cloning of the genome of human spumaretrovirus.

DNA of human spumaretrovirus (HSRV) was cloned from both cDNA and from viral DNA into phage lambda and bacterial plasmid vectors. The recombinant plasmids harboring viral DNA were characterized by Southern blot hybridization and restriction mapping. Physical maps were constructed from cDNA and found to be colinear with the restriction maps obtained from viral DNA. The recombinant clones isolated contained viral DNA inserts which range in size from 2.2 kb to 15.4 kb. The recombinant clones allowed to construct a physical map of the complete HSRV provirus of 12.2 kb.

The nucleotide sequence of the early region of the Tupaia adenovirus DNA corresponding to the oncogenic region E1b of human adenovirus 7.

The nucleotide sequence of the early region E1b of the tree shrew (Tupaia) adenovirus (TAV) DNA has been determined. The sequenced region includes the genes for polypeptides of Mr 15 000, 44 000 and 13 400, which are analogous to the small and large E1b proteins and protein IX, respectively, of the three human adenovirus serotypes 5, 7, and 12. The hexanucleotide consensus signal AATAAA occurs only at the 3' terminus of the gene for protein IX suggesting that the E1 region of TAV encompasses one transcription unit. The amino acid sequences of the TAV polypeptides have a higher degree of homology to those of Ad7 and Ad5 than to those of Ad12.

Identification and mapping of the UL56 gene transcript of herpes simplex virus type 1.

The herpes simplex virus type 1 (HSV-1) strain HFEM is apathogenic for tree shrews and mice by the intraperitoneal application route. This is due to a 4.1 kbp deletion [0.762 to 0.789 map units (mu)] within the BamHI DNA fragment B of the viral genome. With exception of 71 bp the DNA sequences of the deleted region are located within the repetitive DNA sequences of the inverted repeat of the L segment of the HSV-1 genome (IRL). A 1.5 kb RNA hybridizing to the DNA sequences of the HSV-1 genome at map position 0.760-0.762 (BssHII DNA fragment F, part of the BamHI DNA fragment B) was found to be missing in cells infected with HSV-1 HFEM and other apathogenic HSV-1 strains. A detailed analysis of the transcriptional profile of this region of the pathogenic prototype strain HSV-1 F and strand-specific hybridizations revealed that this 1.5 kb RNA species is transcribed at 2 to 4 h p.i. in leftward orientation. The corresponding open reading frame in the HSV-1 genome had been predicted as the UL56 gene. The absence of this 1.5 kb RNA in HSV-1 HFEM-infected cells is due to the fact that the promoter region of the UL56 gene is located within those DNA sequences which are deleted in the HSV-1 HFEM genome. A specific DNA fragment (650 bp) was amplified by reverse polymerase chain reaction using oligonucleotide primers corresponding to the predicted translational start and termination region of the UL56 gene. The corresponding cDNA had been derived from cellular RNA from HSV-1 F-infected cells using oligo(dT) priming. This indicates that the 1.5 kb RNA is the real transcript of the UL56 gene of HSV-1.

Structural organization and analysis of the viral terminase gene locus of Tupaia herpesvirus.

Tupaia herpesvirus (THV) was isolated from spontaneously degenerating tissue cultures of malignant lymphoma, lung, and spleen cell cultures of tree shrews (Tupaia spp.). In order to determine the phylogenetic relatedness of THV the complete nucleotide sequence of the viral terminase (VTER) gene locus (6223 bp) of Tupaia herpesvirus strain 2 (THV-2) was elucidated and analysed. The VTER gene locus, encoding one of the most highly conserved herpes viral proteins is composed of two exons. The intron contains five potential open reading frames (ORFs). The arrangement of these ORFs is colinear with the corresponding regions in the genomes of the mammalian cytomegaloviruses. The precise primary structure of the THV-2 VTER splice junction was determined using RT-PCR and was found to be in agreement with the corresponding splice donor and acceptor sites of the mammalian cytomegaloviruses. The comparison of all six putative THV-2 proteins with the corresponding counterparts in other herpesviruses revealed that THV resides between the Human and the Murine cytomegalovirus (HCMV, MCMV). These results are in agreement with our previous statement, that THV and the known cytomegaloviruses are closely related to each other and should be classified into one taxonomic group. The genetic data presented here and in previous studies are based on the detailed comparison of highly conserved viral genes. Consequently, the classification of the Human and the cytomegaloviruses into the two genera Cyto- and Muromegalovirus, that is mainly based on overall genome structure, should be reconsidered.

Organotropism of latent herpes simplex virus type 1 is correlated to the presence of a 1.5 kb RNA transcript mapped within the BamHI DNA fragment B (0.738 to 0.809 map units).

The transcriptional activity of the DNA sequences within the genome of herpes simplex virus type 1 (HSV-1) at the coordinates 0.760 to 0.762 and their influence in the process of viral latency were investigated. Seven avirulent HSV-1 strains (HFEM, 1752/2, 1752/3, 1752/11, 1469, 1475, 1618), two virulent wild-type HSV-1 strains (F and 17) and three virulent intratypic HSV-1 recombinant viruses (R19, R26, RM1C1) were screened. The virulent HSV-1 strains colonize the ganglia but the avirulent virus strains are only able to persist in the spleen of infected animals (tree shrews). A 1.5 kb RNA transcript was detectable in all virus strains recovered from the ganglia. This RNA transcript hybridised to the HSV-1 DNA sequences at the genome coordinates 0.760 to 0.762 (BssHII DNA fragment F, part of the BamHI DNA fragment B of HSV-1, 0.738 to 0.809 map units (m.u.]. In contrast it was found that the 1.5 kb RNA transcript was missing or its size was changed in cells infected with those HSV-1 strains which were recovered from the spleens of latently infected animals. The state of viral latency of three defined deletion variants of HSV-1 strain 17 (1704, 1705, and 1706) whose genome harbors deletions (2.2 to 5.3 kb) comprising the DNA sequences of the particular region (0.760 to 0.762 m.u.) was investigated. These studies revealed that all three deletion variants could only be recovered from the spleens of latently infected tree shrews.

Restitution of the UL56 gene expression of HSV-1 HFEM led to restoration of virulent phenotype; deletion of the amino acids 217 to 234 of the UL56 protein abrogates the virulent phenotype.

Recently it was shown that the avirulent phenotype of HSV-1 strain HFEM is correlated to the lack of DNA sequences of the promoter region of the UL56 gene. In order to investigate the role of the UL56 gene of HSV-1 in the process of viral pathogenicity in more detail, a complete copy of the UL56 gene of the virulent HSV-1 strain 17 was inserted within the DNA sequences of the incomplete UL56 gene of the genome of HSV-1 strain HFEM. The UL56 gene of HSV-1 strain 17 comprises 1428 bp corresponding to the nucleotide positions (NP) 11,5967-117,395 of the genome of HSV-1 strain 17 (SacII-DNA fragment) containing the promoter region and the entire UL56 gene with identical transcription termination signals. This particular DNA fragment was inserted into the corresponding region of the genome of HSV-1 strain HFEM by co-transfection experiments in which the beta-galactosidase gene served as reporter gene. Those recombinant viruses with the ability to express the UL56 gene were tested for their pathogenicity in vivo. The results of these experiments indicate that the restoration of the viral UL56 gene expression led to the restitution of the virulent phenotype of HSV-1 strain HFEM. The UL56 protein which has been shown to be a component of the virion possesses several characteristic signatures e.g. a hydrophobic domain at the carboxy-terminus between amino acid residues 217 and 234 (VFGVVAIVVVIILVFLWR). In order to investigate the role of this particular signature of the UL56 protein in the process of viral pathogenicity, site-specific mutagenesis was performed for removing the carboxy-terminus of the UL56 protein. The deleted region of the DNA sequences of the UL56 gene between NP 1122-1175 corresponds to NP 116 220-116 373 of the viral genome. The DNA sequences of the UL56 gene of virulent HSV-1 strain 17 and F were replaced by DNA sequences of the truncated UL56 gene by co-transfection experiments in which the beta-galactosidase gene served as a reporter gene. Those recombinant viruses with the ability to express the truncated UL56 gene were examined for their pathogenicity in vivo. The analysis revealed that the expression of the truncated UL56 protein (without hydrophobic domain 217-234 aa) was not sufficient for the maintenance of the virulent phenotype of HSV-1 strains.

In vitro expression of UL56 gene of herpes simplex virus type 1; detection of UL56 gene product in infected cells and in virions.

In order to investigate the functional properties of the UL56 gene of herpes simplex virus type 1 (HSV-1), it was necessary to express the UL56 protein in vitro. The DNA sequences corresponding to the open reading frame of the UL56 gene of HSV-1 strain F were amplified from genomic viral DNA by PCR using primers corresponding to the translational start and termination regions of the UL56 ORF. The PCR product (705 bp) was inserted into the EcoRI/XbaI recognition sites of the bacterial expression vector pMal-c2. This procedure allowed the expression of the viral UL56 gene fused to the maltose-binding protein (MBP) of Escherichia coli, and subsequent cleavage of the fusion protein with the specific protease factor Xa. The induced fusion protein was purified by affinity chromatography using amylose columns. The apparent molecular weight of the fusion protein was about 70 kDa. Factor Xa cleaves the fusion protein into two subfragments of 42 kDa (MBP) and 30 kDa (UL56). Rabbit antisera induced against recombinant UL56 protein were used for detection of the UL56 gene product during the infection cycles of HSV-1. The presence of the UL56 protein was detected in infected cells and in HSV-1 virions by Western blot experiments and by immunofluorescence assays. A strong and increasing cytoplasmic fluorescence was observed in RC-37 cells infected with HSV-1 strain F between 6 and 16 h post-infection. In addition it was found that human HSV-1 IgM/IgG positive convalescent sera recognized the recombinant UL56 protein.

Comparative analysis of the transcripts mapped in the BamHI DNA fragment B of avirulent HSV-1 HFEM, virulent HSV-1 F, and their intratypic recombinant viruses.

HSV-1 HFEM, whose genome harbors a deletion of 4.1 kbp (0.762 to 0.789 map units (mu] is avirulent for mice and tree shrews by the intraperitoneal (i.p.) application route. Insertion of the BamHI DNA fragment B (0.738 to 0.809 mu) and/or the MluI DNA fragment (0.7615 to 0.796 mu) molecularly cloned from virulent HSV-1 F, restored the i.p. pathogenicity to strain HFEM and led to the isolation of virulent intratypic recombinants. In order to determine the RNA transcripts mapped in the BamHI DNA fragment B of the HSV-1 HFEM, HSV-1 F, and their intratypic recombinants R15, R19, R26, and R-Ml-C1, a comparative analysis was performed using Northern blot hybridizations. Two novel RNA transcripts of 3.5 and 1.5 kb were detected which hybridize to the left terminus (0.738 to 0.746 mu) of the BamHI DNA fragment B. The 1.5 kb RNA transcript was missing in the avirulent HSV-1 HFEM. Hybridization with the BssHII DNA fragment F (0.760 to 0.762 mu) led to detection of a 3.5 kb RNA transcript by HSV-1 HFEM which was missing in all other viruses tested. In contrast a 1.5 kb RNA transcript was detectable in all other virus strains with the exception of HSV-1 HFEM. The 3.5 kb transcript hybridized to the right-hand flank of the deleted region in the genome of HSV-1 HFEM (Asp718/SalI DNA fragment; 0.786 to 0.79 mu). The detection of the novel 1.5 kb RNA, which is missing in HSV-1 HFEM, and the appearance of the newly transcribed 3.5 kb RNA in HSV-1 HFEM only, indicates a new open reading frame in this particular region as a consequence of the fusion of the DNA sequences at both ends of the deletion in the genome of HSV-1 HFEM.

Identification, mapping and cloning of the thymidine kinase gene of fish lymphocystis disease virus.

The thymidine kinase (TK) gene of fish lymphocystis disease virus (FLDV) was identified by biochemical transformation of 3T3 TK negative (TK-) to 3T3 TK positive (TK+) cells using specific viral DNA sequences. DNA fragments of the viral genome used in this study were obtained from a defined gene library of FLDV genome containing the complete viral DNA sequences. The selection of the converted cells was carried out under the condition of the HAT selection procedure. The results of these experiments revealed that the EcoRI FLDV DNA fragment C (11.2 kbp; 0.611 to 0.718 map units) is able to transform 3T3 TK- to 3T3 TK+ cells. Additional experiments using the subclones of EcoRI DNA fragment C revealed that DNA sequences of 4.1 kbp size between the coordinates 0.669 to 0.718 of the FLDV genome possessed the ability for biochemical transformation, indicating that the TK gene locus is located in this particular region.

Mutations in the UL53 gene of HSV-1 abolish virus neurovirulence to mice by the intracerebral route of infection.

The cell fusion protein, the product of the UL53 gene, is responsible for intracerebral (IC) pathogenicity of HSV-1. Recombinant HSV-1 R15 is apathogenic to mice by the IC route of inoculation, while intratypic recombinants, in which the UL53 gene in R15 was replaced by an analogous sequence from the pathogenic strain R19, regained IC pathogenicity. The nucleotide sequence of the UL53 gene of HSV-1 strains R15 (apathogenic) and R19 (pathogenic) was determined and compared to that of other pathogenic strains. Four mutations were found which are thought to be responsible for the apathogenic phenotype of HSV-1 strain R15. Northern blot hybridization of RNA extracted from BSC-1 cells infected with several HSV-1 strains indicated that all of the virus strains tested expressed equal amounts of UL53 mRNA in infected cell cultures. Demonstration of the expression of UL53 mRNA in brains of mice infected with HSV-1 strains was made possible by the combined use of a rapid method for mRNA extraction (Oligo dT-linked magnetic beads) and a highly sensitive technique for detection of the existence of the UL53-specific mRNA (cDNA synthesis followed by PCR). It was shown that both pathogenic (KOS and P42) and apathogenic (R15) HSV-1 strains expressed the UL53 gene in brains of IC infected mice.

Determination of the nucleotide sequence flanking the deletion (0.762 to 0.789 map units) in the genome of an intraperitoneally avirulent HSV-1 strain HFEM.

Herpes simplex virus type 1 (HSV-1) strain HFEM which harbours a deletion of 4.1 kbp in its genome (0.762 to 0.789 map units, HpaI DNA fragment P of HSV-1), is apathogenic for mice and tree shrews by the intraperitoneal application route. The exact position of this deletion was determined by DNA sequence analysis. This analysis was performed using the recombinant plasmid pU18HSHF-XmI-B which harbours the flanking genome regions (0.752 to 0.762 and 0.789 to 0.7895 map units) of the deletion in the genome of HSV-1 HFEM, and the recombinant plasmids pU18HSF-XmI-B, pU18HSF-AS, and pHSF-BB-BsH-D, harbouring particular regions of the genome of the virulent HSV-1 strain F at the coordinates 0.752 to 0.761, 0.786 to 0.790, and 0.762 to 0.771, respectively. The comparison of the DNA sequence of this region with the DNA sequences of the corresponding genome regions of the pathogenic HSV-1 strain F and HSV-1 strain 17 showed that the 5' end of the deletion in the genome of HSV-1 HFEM starts at the nucleotide position 3774 of the BamHI DNA fragment B from HSV-1/17. This position is 71 bp upstream of the UL/RL junction of the HSV-1 genome. The 3' terminus of the deletion ends at the nucleotide position 7226 of the BamHI DNA fragment B from HSV-1/17. The position is within the incomplete ninth repetitive box (ACTCC-CACGCACCCCC) and is located 36 bp upstream of the 3' end of the IE 110 mRNA.

Structural organization of a conserved gene cluster of Tupaia herpesvirus encoding the DNA polymerase, glycoprotein B, a probable processing and transport protein, and the major DNA binding protein.

The Tupaia herpesviruses (THVs) have been isolated from malignant lymphoma tissue cultures and from degenerating lung and spleen cell cultures of tree shrews (Tupaia spp.). Recently we succeeded in the localization of the gene locus of the THV DNA polymerase (DPOL) gene within the viral genome. Based on these results the highly conserved gene cluster of herpesviruses encoding the DPOL, the glycoprotein B (gB), a probable processing and transport protein (PRTP), and the major DNA binding protein (DNBI) was characterized in the genome of THV strain 2 (THV-2) in its entirety. The complete nucleotide sequence of the gene cluster was determined and it was discovered that the THV-2 gene products are most closely related to the corresponding proteins of mammalian cytomegaloviruses. The transcriptional activity of the four genes was confirmed by amplification of a part of the corresponding mRNAs obtained from infected cell RNA by RT-PCR. The homology values and the overall structure of the gene cluster, that shows specific colinearity with the corresponding clusters of the mammalian cytomegaloviruses, is further evidence that THV-2 is a member of the subfamily Betaherpesvirinae.

Elimination of UL56 gene by insertion of LacZ cassette between nucleotide position 116030 to 121753 of the herpes simplex virus type 1 genome abrogates intraperitoneal pathogenicity in tree shrews and mice.

In order to investigate whether or not the UL56 gene is involved in those processes determining the viral pathogenicity and latency, a recombinant virus HSV-1-M-LacZ was constructed in which the DNA sequences between nucleotide position (np) 116030 and 121753 were replaced by the E. coli beta-galactosidase (LacZ) gene. This deletion spans from the carboxyterminus of UL55 (np 116030) to the second exon of IE110 (np 121753) eliminating UL56 and the variable region of the BamHI DNA fragment B which were implicated in intraperitoneal pathogenicity and latency. The host range and growth kinetics of the recombinant virus HSV-1 M-LacZ were comparable to the parental strain HSV-1 F. As expected it was found that HSV-1-M-LacZ lost its virulent phenotype and was not able to develop acute infection in animals. The state of the UL56 gene was investigated by determining the cDNA sequence of the UL56 gene transcript of HSV-1 F using PCR products obtained after amplification of the cDNA with oligonucleotide primers corresponding to the translational start and stop codons of this gene. This analysis revealed that the DNA sequence of the UL56 gene of HSV-1 F differed from those DNA sequences determined for the genomic DNA of HSV-1 strain 17. Between nucleotide position 116343 and 116344 two nucleotides -AG- are inserted which prolong the ORF of the UL56 gene to 233 amino acids with a predicted molecular weight of 30 kDa.

Rapid detection of genomic variations in different strains of hantaviruses by polymerase chain reaction techniques and nucleotide sequence analysis.

The polymerase chain reaction (PCR) with subsequent nucleotide sequence analysis was employed to rapidly detect genomic variations among different Hantavirus strains. Using synthetic oligonucleotide primers derived from the M and S segment RNAs of nephropathia epidemica virus strain Hällnäs B1 (NEV) we succeeded in amplifying the corresponding sequences of Hantaan and Puumala viruses. The nucleotide sequences of the cDNAs derived from the Puumala M and S RNA segments were analyzed. It was found that the particular nucleotide sequences of Puumala M and S segments were 81% and 82% homologous to the corresponding genomic segments of NEV, respectively. The amino acid homology was 94% for both segments. In contrast, the degree of homology to the corresponding Hantaan M and S genomic RNA segments was 63% at the nucleotide level for both segments and 53 and 55% at the deduced amino acid level, respectively. This demonstrates that Puumala virus is very similar to NEV and significantly different from Hantaan virus at both the nucleotide and protein level.

Determination of the coding capacity of the BamHI DNA fragment B of apathogenic Herpes simplex virus type 1 strain HFEM by DNA nucleotide sequence analysis.

Herpes simplex virus type 1 (HSV-1) strain HFEM acquired an apathogenic phenotype due to a deletion within the DNA sequences of the BamHI DNA fragment B of the viral genome. In order to investigate the coding strategy of this particular region of the genome of HSV-1 strain HFEM the DNA nucleotide sequence of the BamHI DNA fragment B was determined. This analysis revealed that the BamHI DNA fragment B of HSV-1 strain HFEM comprises 6593 bp, corresponding to the nucleotide positions (np) 113322 to 117088 and np 120643 to 123465 of the genome of HSV-1 strain 17. According to these data the deletion of the genome of HSV-1 strain HFEM occurred between the np 117089 and 120642. The promoter region of the UL56 gene of HSV-1 strain HFEM is a part of the deleted DNA sequences. Therefore, this gene of HSV-1 strain HFEM is affected and cannot be expressed. The first 35 amino acid (AA) residues of the deduced amino acid sequence of the UL56 open reading frame (ORF) were found to be identical to the amino acid sequence of the UL56 genes of HSV-1 strains 17 and F. However, due to a deletion at np 3494 of the BamHI DNA fragment B of HSV-1 strain HFEM the amino acid composition of the predicted UL56 gene of HSV-1 strain HFEM is different from HSV-1 strain 17 between amino acid positions 36 and 233. In addition the deduced amino acid sequence of the IRL (inverted repeat of the long segment) copy of the IE110 gene of HSV-1 strain HFEM was found to be about 342 amino acids shorter than the amino acid sequence of IE110 gene of HSV-1 strain 17 (775 AA). This was based on a point mutation which was detected within the DNA sequences of Exon 3 of this copy of IE110 gene of HSV-1 strain HFEM.

Antigenicity of hantavirus nucleocapsid proteins expressed in E. coli.

DNA clones representing the small genomic segment of Nephropathia epidemica virus strain Hällnäs B1 (NEV) and Hantaan virus strain 76-118 (HTV) encoding their nucleocapsid proteins were inserted into the E. coli vector pIN-III-ompA for secretion of proteins into the periplasmic space. The complete HTV and NEV nucleocapsid proteins and two truncated versions of the NEV nucleocapsid proteins were expressed as fusion proteins. Unexpectedly, all products accumulated as insoluble aggregates. Most of the ompA signal peptide remained uncleaved. However, nucleocapsid fusion proteins could be purified from the insoluble fraction by extraction with 8 M urea followed by separation on SDS-PAGE and electroelution. Rabbits were immunized with the eluted proteins and the resulting antibodies reacted specifically with authentic viral nucleocapsid proteins of HTV and NEV. The recombinant nucleocapsid proteins were found to react specifically with various hantavirus-immune sera, but not with human control sera, indicating their suitability as potential diagnostic antigens. This is the first report on the expression of a protein of a NEV serotype strain of hantaviruses by use of recombinant DNA techniques.

Replacement of the deletion in the genome (0.762-0.789 mu) of avirulent HSV-1 HFEM using cloned MluI DNA fragment (0.7615-0.796 mu) of virulent HSV-1 F leads to generation of virulent intratypic recombinant.

The HFEM strain of HSV-1 is apathogenic for the tree shrew by the intraperitoneal (i.p.) route because of a deletion in the genome coordinates 0.762-0.789. Insertion of the MluI DNA fragment (coordinates 0.7615-0.789) cloned from HSV-1 strain F, which is pathogenic for the tree shrew, restored the i.p. pathogenicity to strain HFEM. The recombinant designated R-M1-C1 was highly pathogenic for the tree shrew, but slightly virulent for inbred mouse strain A. It thus appears that the viral DNA sequence involved in the i.p. pathogenicity of HSV-1 is located within the genome coordinates 0.761-0.796. This sequence is recognized differently by the cellular elements involved in HSV-1 infection in the tree shrew and the mouse.