DNA Sequence Anaylysis of an Immediate-Early Gene Region of the Herpes Simplex Virus Type 1 Genome (Map Coordinates 0.950 to 0.978)

Abstract

SUMMARYThe nucleotide sequence of 4 kilobases of DNA from within the short region of the genome of herpes simplex virus type 1 has been determined. This portion of DNA contains the junctions of the terminal and internal inverted repeat sequence components with the unique sequence component and the 5' regions of the genes which encode the Vmw 12, Vmw 68 and Vmw 175 immediate-early polypeptides. The transcription and translation initiation sites of all three genes and the 5' and 3' boundaries of the Vmw 12 and Vmw 68 gene introns have been localized on the DNA sequence and shown to be flanked by sequences which resemble those found in similar positions in other eukaryotic genes. For the Vmw 12 and Vmw 68 genes the promoters, the 5'-non-coding regions of the mRNAs, and the introns lie within the terminal and internal inverted repeats respectively while the polypeptide-coding regions lie in the short unique component. The introns consist largely of tandemly reiterated copies of a 22-nucleotide sequence: the Vmw 12 gene intron contains seven of these and the Vmw 68 gene intron five. The Vmw 175 gene is located entirely within the short repeats, of which those regions sequenced here have the extremely high G + C content of 78 %, in marked contrast to the value of 66 % obtained for the adjacent region of the unique sequence component. Prediction of the complete amino acid sequence of the Vmw 12 polypeptide, accounting for a mol. wt. of 9830, and of the first 523 amino-terminal amino acids of the Vmw 175 polypeptide has been made from the DNA sequence. The polypeptide-coding region of the Vmw 175 gene has an average G + C content of 80% but nevertheless specifies a wide range of amino acid types because of the maximal assignment of G and C residues to codon third base positions.

A Herpes Simplex Virus Type 1 Mutant with a Deletion in the Polypeptide-coding Sequences of the ICP4 Gene

“…From the sequence data of Murchie & McGeoch (1982) it can be concluded that any mutation affecting the 1844 base pair BamHI Y fragment 0000-6503 © 1985 SGM et al, 1979). However, the lesion could still reside within the 275 base pairs of the 5" non-coding region of the gene (Murchie & McGeoch, 1982) which are also part of fragment Y. In order to carry out a fine mapping of the mutation in B a m H I Y, both the w.t.…”

“…Autoradiographs of lanes (a) determined were residues 570 to 713 for the w.t. type and residues 570 to 786 for the mutant; this numeration takes as number 1 the first residue of the presumed translation initiator codon (Murchie & McGeoch, 1982). The sequences presented in Fig.…”

“…2a). The sequence data obtained by Murchie & McGeoch (1982) (Towbin et al, 1979) and exposed to the 58S antibody (Showalter et al, 1981 ). Bound 58S antibody was detected by peroxidase-conjugated goat antimouse antibody.…”

Identification of Two Herpes Simplex Virus Type 1-induced Proteins (21K and 22K) which Interact Specifically with the a Sequence of Herpes Simplex Virus DNA

“…Plasmid pGX60 contains part of the sequences encoding the immediate-early polypeptide IE 175. This region of HSV DNA is not known to contain control elements (Murchie & McGeoch, 1982).…”

“…Both are inserted between the BamHl and Sail sites of pAT153 and can be excised using a combination of these two enzymes, pGX60 was produced by inserting BamHI y (1840 bp) into the BamHI site of pAT153 (Stow, 1982). Digestion with HinfI yields a 619 bp fragment (Murchie & McGeoch, 1982). The derivations of plasmids pG3b, pR 10 and pR9 which have a sequence containing fragments have all been described .…”