Anatomy of herpes simplex virus DNA: strain differences and heterogeneity in the locations of restriction endonuclease cleavage sites.

Abstract

We are attempting to construct maps of restriction enzyme fragments from the DNA of herpes simplex virus (HSV), and wish to report on some unusual observations relating to the structure of HSV DNA molecules that emerged from these studies. We have also been concerned about possible genetic differences between the various strains and isolates of HSV-1 and HSV-2. Two clinical isolates, designated HSV-1(Fl) and HSV-2(G1), have been used in this laboratory as prototypes of naturally occurring strains and have been passaged at low multiplicity a maximum of four times in HEp-2 (human epidermoid carcinoma) cells to avoid selection of variants in culture. However, established "laboratory" strains characterized by histories of numerous passages outside the human (5), and show apl)roximately 50% homology with 85% matching of base pairs (8). (iv) An additional DNA species with unusually high buoyant density (>1.731 g/cm3) may also be present in viral l)reparations after serial passages at high multiplicity (9). MATERIALS AND METHODSVirus Strains. HSV-1 strains, designated Fl, F5, and F9, andl HSV-2(G1) are independent isolates from patients at the University of Chicago Hospitals (1). HSV-1 (A428) was isolated from human trigeminal ganglion by Dr. A. Nahmias, Atlanta, Ga. All isolates were passaged a maximum of four times at low multiplicity inlHEp-2 cells. Despite these immunological tests, l)r. F. Rapp found in 1973 that the DNA of the Justin strain had a buoyant density pattern similar to that of HSV-2 (personal communication, A. Sabin).We believe that this was probably due to the fact that most of the Justin strain viral D)NA is defective-as much as 80% in some of our preparations. In our laboratory, the Justin strain was identified as HSV-l on the basis of structural polypeptide content and by its D)NA buoyant density and restriction endonuclease cleavage pattern. 1768

One Functional Copy of the Long Terminal Repeat Gene Specifying the Immediate-early Polypeptide IE 110 Suffices for a Productive Infection of Human Foetal Lung Cells by Herpes Simplex Virus

“…Our observations raise the question why all field isolates examined to date are diploid for repetitive sequences (Hayward et al, 1975 a;Skare et al, 1975;Lonsdale et al, 1979). We can envisage several explanations.…”

“…4. Those endonucleases which do not cleave both pairs of repetitions generate half-and quarter:molar bands in restriction profiles (Hayward et al, 1975 b;Clements et al, 1976;Wilkie & Cortini, 1976). These bands represent restriction fragments mapping at the termini and the joint between L and S, and result from the presence of equal amounts of the four genome isomers within a DNA population.…”

“…J. Davison & N. M. Wilkie, unpublished results). The L and S segments usually invert freely about the joint, resulting in equal amounts of both orientations of each segment in virion DNA (Hayward et al, 1975 b;Clements et aL, 1976: Delius & Clements, 1976.…”

Variable Restriction Endonuclease Sites of Herpes Simplex Virus Type 1 Isolates from Encephalitic, Facial and Genital Lesions and Ganglia

“…The questions arise whether HSV-1 substrains exist with special predilection for particular anatomical regions and whether there is evidence for geographical substrain clustering. Whilst HSV-1 intra-strain heterogeneity is well established (Heine et al, 1974;Skate et al, 1975;Cassai et al, 1975;Hayward et al, 1975), data suggesting non-random association of HSV-1 isolate subgroups with either anatomical sites or geographical regions are very limited (Pereira et al, 1976;Chaney et al, 1983). As strains of HSV-1 differ considerably in pathogenicity for mice (Hill et al, 1975 ;Kiimel et al, 1982), the possibility cannot be ruled out that there are virus substrain-specific factors involved in herpes encephalitis, in latency and in other herpetic infections (Baringer, 1975;Nahmias et al, 1982).…”

Analysis of herpes simplex virus type 1 restriction fragment length polymorphism variants associated with herpes gladiatorum and Kaposi's varicelliform eruption in sumo wrestlers

“…Complement (0.25 units) was added to each well of 96-well micro-test plates for CRN antibody titrations (Yoshino & Abe, 1981). RFLP analyses were performed as previously described Ozawa et al, 2006) using the restriction endonuclease maps of strain F as references (Hayward et al, 1975;Locker & Frenkel, 1979;Roizman, 1979; Roizman & Tognon, 1983). The terminal internal and 'joint' sequences containing the 280 bp repeat, consisting predominantly of the terminal reiterated sequence 'a' (Davison & Wilkie, 1981;Locker & Frenkel, 1979;Roizman, 1979) (Fig.…”