Identification of the Marek’s Disease Virus Serotype 2 Genes Homologous to the Glycoprotein B (UL27), ICP18.5 (UL28) and Major DNA-Binding Protein (UL29) Genes of Herpes Simplex Virus Type 1

Abstract

ABSTRACT. We determined the nucleotide sequence of non-pathogenic Marek's disease virus serotype 2 (MDV2) strain HPRS24 glycoprotein B (gB) (UL27), ICP18.5 (UL28) and major DNA-binding protein (MDBP) (UL29) genes homologous to herpes simplex virus type 1 (HSV-1). The sequence data revealed that important motives in the proteins are conserved in MDV2 ICP18.5 and MDBP, however the sequence of viral DNA replication origin which exists in the regions between the UL29 and UL30 genes of other alphaherpesviruses was not found in the regions of the MDV2 genome. By northern blot analyses, we also demonstrated that 8.9, 5.0 and 2.6 kb transcripts were actually transcribed from the sequenced region in MDV2-infected cells. 1B) were then deleted by exonuclease III to construct deletion clones. After that DNA sequences were determined on both strands using an autosequencer system (Applied Biosystems, Foster City, CA). DNA and amino acid sequences were analyzed with the computer program GENETYX-MAC (version 8.0). Homology searches and multiple sequence alignments from the GenBank and EMBL Data Libraries were performed using the UWGCG programs BESTFIT and PILEUP, respectively. The nucleotide sequences of the MDV2 UL27, UL28 and UL29 homologues have been submitted to the DDBJ, EMBL and GenBank Databanks with the Accession No. AB024711.The location of the MDV2 (HPRS24 strain) UL27, UL28 and UL29 homologues in virus genome, pertinent restriction endonuclease sites are illustrated in Fig. 1. We determined the nucleotide sequence of a total of 9,650 bp containing the putative sequences of MDV2 UL27, UL28 and UL29 homologues of the HSV-1 gene (Fig. 1C).To find out whether transcripts are really transcribed from the domain of MDV2 genome containing the identified genes of MDV2 UL27, UL28, and UL29, northern blot analyses were performed using the different probes. The maintenance of primary chicken embryo fibroblasts (CEF) and infection of CEF with MDV2 strain HPRS24 were performed as described previously [14]. To isolate total RNA, monolayers were harvested at 96 hr postinfection and treated with 0.05% ethylenediaminetetraacetic acid in phosphate-buffered saline. Total RNAs were extracted from these lysates by ISOGEN (Nippon Gene, Tokyo, Japan). Total RNA was separated on an agarose formaldehyde-gel Marek's disease virus (MDV) is a cell-associated alphaherpesvirus of poultry and causes a naturally occurring malignant T cell lymphoma in chickens. The virus is divided into three serotypes.They belong to alphaherpesviruses, based on their genomic structures and arrangements [4,11,21]. The disease can be prevented by vaccination with attenuated MDV serotype 1 (MDV1), nonpathogenic MDV serotype 2 (MDV2) or herpesvirus of turkeys (HVT) known as a MDV serotype 3 (MDV3). The MDV2 genome differs in restriction endonuclease digestion patterns from those of either MDV1 or HVT [10,25,30]. However, information about the genomic analysis of MDV2 is limited and only the genes encoding for glycoproteins gB (SB-1 strain) [34] [22] have been reported....

The complete unique long sequence and the overall genomic organization of the GA strain of Marek's disease virus

“…cDNA sequencing has revealed that the v-LIP transcript is made up of two exons; a signal sequence is conferred from a very short first exon and the catalytic domain is encoded by the much longer second exon. This pattern is conserved in the close homolog present in MDV-2 HPRS-24 (22). Remarkably, aside from the MDV-2 homolog, the closest homologs to v-LIP are ORFs from several avian and fowl adenoviruses (GenBank accession nos.…”

“…Both the repeat and the UL͞RL junction regions also show significant divergence among the different serotypes of MDV, underscoring the potential importance of these MDV-specific ORFs in conferring the oncogenicity of the viruses (GenBank accession no. AB024414) (21,22). In the ensuing sections, we will first describe MDV genes that have homologs in HSV-1, followed by a more detailed discussion of MDV-specific ORFs.…”

Molecular characterization of the herpes simplex virus 1 (HSV-1) homologues, UL25 to UL30, in duck enteritis virus (DEV)

“…It was shown that the UL28 gene product in HSV-1, together with those from UL6, UL25, UL32 and UL33, are required for full capsid formation, viral DNA packaging, and viral glycoprotein expression on the surface of virus-infected cells (Addison et al, 1990;Tengelsen et al, 1993). Alignment analysis indicated that the predicted nuclear localization sequence, RRK, of UL28 protein in DEV clone-03 was conserved among the herpesviruses (Kato et al, 1999), which suggested that DEV UL28 protein may be related to the capsidation of virus.…”

Genomic Variation of the Fibropapilloma-Associated Marine Turtle Herpesvirus across Seven Geographic Areas and Three Host Species

“…The predicted amino acid sequence of the ORF at 13735 to 10877 bears herpesvirus single-stranded DNA binding protein homology, with 29 to 34% identity and 46 to 51% similarity to the model alphaherpesviruses (Table 2). This homology includes a putative conserved zinc finger motif at predicted positions 491 to 502 (CDLCDVETRHVC; Zn finger consensus sequence is C-X 2-5 -C-X 2-15 -C/H-X 2-4 -C/H, where bold indicates conserved residues) (11), followed by a putative conserved DNA binding domain (R 795 AEKVMQLQPILNGPHGYLLKRFHERLFPTN KAIS AMAFWNRAQNNK; the U L 29 DNA binding consensus motif is K/R-X 2-3 -K/R-X 9-10 -F-X 4 -F-X 1-3 -K/R-X 10-12 -W-X 6 -R (16). The U L 29 nucleotide sequence reported here is Ͼ99% identical to a FPTHV U L 29 sequence in GenBank (27).…”