1 Department of Virology, Faculty of Medicine, Kyushu University, Fukuoka 812-8582, Japan
2 Department of Obstetrics and Gynecology, Mizonokuchi Hospital, Teikyo University, Kawasaki 213-8507, Japan
Correspondence
Kenichi Umene
umene{at}virology.med.kyushu-u.ac.jp
![]() |
ABSTRACT |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
![]() |
INTRODUCTION |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
The genome of HSV-1 is a 152 kb linear duplex DNA molecule (McGeoch et al., 1988) (Fig. 1
a). Epidemiologically unrelated HSV-1 strains can usually be differentiated by analysing variations in restriction endonuclease (RE) cleavage patterns (Buchman et al., 1978
, 1980
; Chaney et al., 1983
; Lonsdale et al., 1980
). Variations in RE cleavage patterns between HSV-1 strains are divided into two types (Umene et al., 1984
; Umene & Yoshida, 1989
, 1993
, 1994
; Umene, 1998a
, b
). One type of variation, termed restriction fragment length polymorphism (RFLP), is due mostly to a gain or loss of an RE cleavage site and causes a simple change in RE cleavage patterns. RFLPs are stable and serve as physical markers of the HSV-1 genome in molecular epidemiological and evolutionary studies. Another type of variation appears as irregularities in RE-cleaved fragments derived from certain regions of the HSV-1 genome. This variation was found in all strains analysed and was termed common type (Umene et al., 1984
; Umene, 1998a
, b
). Common type variation is located in fragments containing tandemly repeated sequences and is due to variation in copy number or nucleotide sequence of the reiterations (Davison & Wilkie, 1981
; Mocarski & Roizman, 1981
; Rixon et al., 1984
; Umene, 1985b
, 1991
, 1998a
, b
, 1999
). However, use of common type variations to distinguish HSV-1 strains has been avoided as they may be too unstable to qualify as markers.
|
An HSV-1 strain (ancestor) is presumed to go through variations and consequently diverge into distinguishable strains (descendants) (McGeoch & Cook 1994, McGeoch et al., 1995
; Sakaoka et al., 1994
; Umene & Sakaoka, 1997
, 1999
). Shortly after the generation of descendant viruses (the relatively earlier stage of divergence), a significant difference between descendant isolates is not usually detectable as either RFLP or common type variation (Fig. 2
a, b). Long after the generation of descendant viruses (the relatively later stage of divergence), significant difference is detectable as both RFLP and common type variation (Fig. 2a, b
). A state of transition from the relatively earlier stage to the relatively later stage must have been present during divergence. Common type variation occurs at a faster pace than nucleotide substitution detectable by RFLP patterns, thereby allowing a finer level of resolution of the diversification process. Thus, the presence of a state of transition was considered to be identifiable if a significant difference could be detected as common type variation between HSV-1 isolates from the same patient that could not be distinguished in terms of RFLP (Fig. 2
). In the present study, variation was analysed between HSV-1 isolates which were isolated from genital lesions of the same individual at different episodes of recurrence. Significant differences between these HSV-1 isolates were detected as common type variation but not as RFLP. Thus, the state of transition from relatively earlier stage to relatively later stage was shown to exist.
|
![]() |
METHODS |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
|
Gel electrophoresis and Southern blot hybridization.
DNA digested with an RE was separated in a 0·8 % (w/v) agarose gel and in a 5 % (w/v) acrylamide gel, as described by Umene (1985b). Southern blot hybridization was carried out on a Biodyne B membrane (Pall Biosupport), according to the manufacturer's instructions. DNA fragments used as probes were prepared from hybrid phages and plasmids and were labelled with [
-32P]dCTP, as described by Umene & Yoshida (1989)
.
![]() |
RESULTS |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
In the present study, we analysed the RFLPs of 16 HSV-1 isolates in six sets (Table 1) using three 4-bp REs (HaeIII, HhaI and MboI) in the same manner as in previous studies (Umene, 1987
; Umene & Sakaoka, 1991
; Umene & Yoshida, 1994
). The RFLP patterns (using three 4-bp REs) of HSV-1 strains from each set were the same. Thus, HSV-1 isolates belonging to each set were not differentiated by RFLPs using 6-bp and 4-bp REs.
Analyses of variation of reiterated sequences in each of six sets of HSV-1 isolates
Variation of reiterations I, IV, and VII was analysed in six sets of HSV-1 isolates.
Reiteration VII.
Southern blot hybridization profiles of reiteration VII are shown in Fig. 3. Regions containing reiteration VII in isolates of sets 14 and 6 were detected as a 0·17 kb fragment (Fig. 3a
, lanes 110 and Fig. 3b
, lanes 1, 2, 5 and 6) and those of set 5 as a 0·15 kb fragment (Fig. 3b
, lanes 3 and 4). Therefore, HSV-1 isolates in each set were not differentiated by reiteration VII.
|
|
|
![]() |
DISCUSSION |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
Differences in the degree of stability between reiterations in the HSV-1 genome
Differences in reiteration VII were not detected between HSV-1 isolates from the same patient yet differences were evident in reiterations I and IV (Figs 35). Thus, reiteration VII is assumed to be the most stable (Umene & Yoshida, 1989
).
In our previous study, differences in reiteration IV were not detected between single-plaque isolates and between clinical isolates from the same patient (Umene & Yoshida, 1989). However, significant differences in reiteration IV were detected in the present study between clinical isolates, which were isolated from the same patient and could not be distinguished in terms of RFLP variation (sets 1 and 2) (Fig. 4a
, lanes 14). Differences in reiteration IV were detected in two of the three multiple-recurrence sets but in none of the primary-recurrence sets; hence, the probability of detection of events relating to differences in reiteration IV seemed to be larger in multiple-recurrence sets than in primary-recurrence sets.
Reiteration I was considered to be capable of serving as a marker for differentiating HSV-1 strains, since the major (or single) fragment containing reiteration I was the same size in single-plaque isolates (Umene, 1991; Umene & Yoshida, 1989
). Differences in the major fragment of reiteration I between clinical isolates from the same patient were detected in multiple-recurrence sets 1 and 3, but not in primary-recurrence sets (Fig. 5
). Maertzdorf et al. (1999)
did not adopt reiteration I to differentiate HSV-1 strains, because PCR-amplified fragments containing reiteration I were not identical in size between single-plaque isolates. Reiteration I is presumed to be unsuitable for PCR amplification since it has a high G+C content, a strong bias towards purines on one strand and pyrimidines on the other, and adopts unusual DNA structures (Martin & Weber, 1998
; Wells et al., 1988
; Wohlrab et al., 1987
). However, reiteration I can serve as marker if HSV-1 DNA is analysed without PCR amplification, although it appears to be less stable than reiterations IV and VII.
Development of HSV-1 lineages within the body of an individual
The probability of detecting differences between progeny viruses is assumed to increase in proportion to the number of rounds of virus DNA replication. Isolates of the multiple-recurrence sets are presumed to have passed through more rounds of DNA replication than strains of the primary-recurrence sets. Thus, differences between isolates from the same patient would be expected to be more frequent in multiple-recurrence sets than in primary-recurrence sets. As expected, differences were detected in each multiple-recurrence set but not in primary-recurrence sets. This supports the view that a significantly larger number of replicative cycles separates the viruses shed in different recurrences of the same primary infection (multiple-recurrence set) than occurs between primary infection and the first recurrence (primary-recurrence set). Through a significantly larger number of replicative cycles occurring during long term infection, HSV-1 is assumed to pass into a state of transition from a relatively earlier stage (Fig. 2b). Isolates of set 3 are possibly less differentiated than isolates of sets 1 and 2 because of a common sharing of ladder-like fragments of reiteration I and lack of differences in reiteration IV (Fig. 4a
, lanes 510 and Fig. 5a
, lanes 38). Reiterated sequences are useful for study of the epidemiology and evolution of HSV-1.
![]() |
ACKNOWLEDGEMENTS |
---|
![]() |
REFERENCES |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
Buchman, T. G., Simpson, T., Nosal, C., Roizman, B. & Nahmias, A. J. (1980). The structure of herpes simplex virus DNA and its application to molecular epidemiology. Ann N Y Acad Sci 354, 279290.[Medline]
Chaney, S. M. J., Warren, K. G., Kettyls, J., Zbitnue, A. & Subak-Sharpe, J. H. (1983). A comparative analysis of restriction enzyme digests of the DNA of herpes simplex virus isolated from genital and facial lesions. J Gen Virol 64, 357371.[Abstract]
Davison, A. J. & Wilkie, N. M. (1981). Nucleotide sequences of the joint between the L and S segments of herpes simplex virus types 1 and 2. J Gen Virol 55, 315331.[Abstract]
Dolan, A., Jamieson, F. E., Cunningham, C., Barnett, B. C. & McGeoch, D. J. (1998). The genome sequence of herpes simplex virus type 2. J Virol 72, 20102021.
Everett, R. D. (2000). ICP0, a regulator of herpes simplex virus during lytic and latent infection. Bioessays 22, 761770.[CrossRef][Medline]
Hashido, M., Inouye, S. & Kawana, T. (1997). Differentiation of primary from nonprimary genital herpes infections by a herpes simplex virus-specific immunoglobulin G avidity assay. J Clin Microbiol 35, 17661768.[Abstract]
Hashido, M., Lee, F. K., Nahmias, A. J., Tsugami, H., Isomura, S., Nagata, Y., Sonoda, S. & Kawana, T. (1998). An epidemiologic study of herpes simplex virus type 1 and 2 infection in Japan based on type-specific serological assays. Epidemiol Infect 120, 179186.[CrossRef][Medline]
Kawana, T., Kawagoe, K., Takizawa, K., Chen, T., Kawaguchi, T. & Sakamoto, S. (1982). Clinical and virologic studies on female genital herpes. Obstet Gynecol 60, 456461.[Abstract]
Kinghorn, G. R. (1993). Genital herpes: natural history and treatment of acute episodes. J Med Virol Supplement 1, 3338.
Lafferty, W. E., Coombs, R. W., Benedetti, J., Critchlow, C. & Corey, L. (1987). Recurrences after oral and genital herpes simplex virus infection: influence of site of infection and viral type. N Engl J Med 316, 14441449.[Abstract]
Lafferty, W. E., Downey, L., Celum, C. & Wald, A. (2000). Herpes simplex virus type 1 as a cause of genital herpes: impact on surveillance and prevention. J Infect Dis 181, 14541457.[CrossRef][Medline]
Lonsdale, D. M., Brown, S. M., Lang, J., Subak-Sharpe, J. H., Koprowski, H. & Warren, K. G. (1980). Variations in herpes simplex virus isolated from human ganglia and a study of clonal variation in HSV-1. Ann N Y Acad Sci 354, 291308.[Abstract]
McGeoch, D. J. & Cook, S. (1994). Molecular phylogeny of the alphaherpesvirinae subfamily and a proposed evolutionary timescale. J Mol Biol 238, 922.[CrossRef][Medline]
McGeoch, D. J., Dalrymple, M. A., Davison, A. J., Dolan, A., Frame, M. C., McNab, D., Perry, L. J., Scott, J. E. & Taylor, P. (1988). The complete DNA sequence of the long unique region in the genome of herpes simplex virus type 1. J Gen Virol 69, 15311574.[Abstract]
McGeoch, D. J., Cook, S., Dolan, A., Jamieson, F. E. & Telford, E. A. R. (1995). Molecular phylogeny and evolutionary timescale for the family of mammalian herpesviruses. J Mol Biol 247, 443458.[CrossRef][Medline]
Maertzdorf, J., Remeijer, L., Van Der Lelij, A., Buitenwerf, J., Niesters, H. G. M., Osterhaus, A. D. M. E. & Verjans, G. M. G. M. (1999). Amplification of reiterated sequences of herpes simplex virus type 1 (HSV-1) genome to discriminate between clinical HSV-1 isolates. J Clin Microbiol 37, 35183523.
Maertzdorf, J., van der Lelij, A., Baarsma, G. S., Osterhaus, A. D. M. E. & Verjans, G. M. G. M. (2000). Herpes simplex virus type 1 (HSV-1)-induced retinitis following herpes simplex encephalitis: indications for brain-to-eye transmission of HSV-1. Ann Neurol 48, 936939.[CrossRef][Medline]
Martin, D. W. & Weber, P. C. (1998). Role of the DR2 repeat array in the regulation of the ICP34.5 gene promoter of herpes simplex virus type 1 during productive infection. J Gen Virol 79, 517523.[Abstract]
Mertz, G. J. (1990). Genital herpes simplex virus infections. Med Clin N Am 74, 14331454.[Medline]
Mocarski, E. S. & Roizman, B. (1981). Site-specific inversion sequence of the herpes simplex virus genome: domain and structural features. Proc Natl Acad Sci U S A 78, 70477051.[Abstract]
Mocarski, E. S., Deiss, L. P. & Frenkel, N. (1985). Nucleotide sequence and structural features of a novel US-a junction present in a defective herpes simplex virus genome. J Virol 55, 140146.[Medline]
Preston, C. M. (2000). Repression of viral transcription during herpes simplex virus latency. J Gen Virol 81, 119.
Remeijer, L., Maertzdorf, J., Doornenbal, P., Verjans, G. M. G. M. & Osterhaus, A. D. M. E. (2001). Herpes simplex virus 1 transmission through corneal transplantation. Lancet 357, 442.[CrossRef][Medline]
Remeijer, L., Maertzdorf, J., Buitenwerf, J., Osterhaus, A. D. M. E. & Verjans, G. M. G. M. (2002). Corneal herpes simplex virus type 1 superinfection in patients with recrudescent herpetic keratitis. Investig Ophthalmol Vis Sci 43, 358363.
Ribes, J. A., Steele, A. D., Seabolt, J. P. & Baker, D. J. (2001). Six-year study of the incidence of herpes in genital and nongenital cultures in a central Kentucky Medical Center patient population. J Clin Microbiol 39, 33213325.
Rixon, F. J., Campbell, M. E. & Clements, B. (1984). A tandemly reiterated DNA sequence in the long repeated region of herpes simplex virus type 1 found in close proximity to immediate-early mRNA 1. J Virol 52, 715718.[Medline]
Sakaoka, H., Kurita, K., Iida, Y., Takada, S., Umene, K., Kim, Y. T., Ren, C. S. & Nahmias, A. J. (1994). Quantitative analysis of genomic polymorphism of herpes simplex virus type 1 strains from six countries: studies of molecular evolution and molecular epidemiology of the virus. J Gen Virol 75, 513527.[Abstract]
Stanberry, L., Cunningham, A., Mertz, G. & 7 other authors. (1999). New developments in the epidemiology, natural history and management of genital herpes. Antiviral Res 42, 114.[CrossRef][Medline]
Sucato, G., Wald, A., Wakabayashi, E., Vieria, J. & Corey, L. (1998). Evidence of latency and reactivation of both herpes simplex virus (HSV)-1 and HSV-2 in the genital region. J Infect Dis 177, 10691072.[Medline]
Taylor, S., Drake, S. & Pillay, D. (1999). Genital herpes, the new paradigm. J Clin Pathol 52, 14.
Umene, K. (1985a). Intermolecular recombination of the herpes simplex virus type 1 genome analysed using two strains differing in restriction enzyme cleavage sites. J Gen Virol 66, 26592670.[Abstract]
Umene, K. (1985b). Variability of the region of the herpes simplex virus type 1 genome yielding defective DNA: SmaI fragment polymorphism. Intervirology 23, 131139.[Medline]
Umene, K. (1987). Restriction endonucleases recognizing DNA sequences of four base pairs facilitate differentiation of herpes simplex virus type 1 strains. Arch Virol 97, 197214.[Medline]
Umene, K. (1991). Recombination of the internal direct repeat element DR2 responsible for the fluidity of the a sequence of herpes simplex virus type 1. J Virol 65, 54105416.[Medline]
Umene, K. (1998a). Herpesvirus: Genetic Variability and Recombination. Fukuoka: Touka Shobo.
Umene, K. (1998b). Molecular epidemiology of herpes simplex virus type 1. Rev Med Microbiol 9, 217224.
Umene, K. (1999). Mechanism and application of genetic recombination in herpesviruses. Rev Med Virol 9, 171182.[CrossRef][Medline]
Umene, K. (2001). Cleavage in and around the DR1 element of the a sequence of herpes simplex virus type 1 relevant to the excision of DNA fragments with length corresponding to one and two units of the a sequence. J Virol 75, 58705878.
Umene, K. & Enquist, L. W. (1981). A deletion analysis of lambda hybrid phage carrying the US region of herpes simplex virus type 1 (Patton). I. Isolation of deletion derivatives and identification of chi-like sequences. Gene 13, 251268.[CrossRef][Medline]
Umene, K. & Yoshida, M. (1989). Reiterated sequences of herpes simplex virus type 1 (HSV-1) genome can serve as physical markers for the differentiation of HSV-1 strains. Arch Virol 106, 281299.[Medline]
Umene, K. & Sakaoka, H. (1991). Homogeneity and diversity of genome polymorphism in a set of herpes simplex virus type 1 strains classified as the same genotypic group. Arch Virol 119, 5365.[Medline]
Umene, K. & Yoshida, M. (1993). Genomic characterization of two predominant genotypes of herpes simplex virus type 1. Arch Virol 131, 2946.[Medline]
Umene, K. & Yoshida, M. (1994). Preparation of herpes simplex virus type 1 genomic markers to differentiate strains of predominant genotypes. Arch Virol 138, 5569.[Medline]
Umene, K. & Sakaoka, H. (1997). Populations of two Eastern countries of Japan and Korea and with a related history share a predominant genotype of herpes simplex virus type 1. Arch Virol 142, 19531961.[CrossRef][Medline]
Umene, K. & Sakaoka, H. (1999). Evolution of herpes simplex virus type 1 under herpesviral evolutionary processes. Arch Virol 144, 637656.[CrossRef][Medline]
Umene, K. & Kawana, T. (2000). Molecular epidemiology of herpes simplex virus type 1 genital infection in association with clinical manifestations. Arch Virol 145, 505522.[CrossRef][Medline]
Umene, K., Eto, T., Mori, R., Takagi, Y. & Enquist, L. W. (1984). Herpes simplex virus type 1 restriction fragment polymorphism determined using Southern hybridization. Arch Virol 80, 275290.[Medline]
Vanderhooft, S. & Kirby, P. (1992). Genital herpes simplex virus infection: natural history. Semin Dermatol 11, 190199.[Medline]
Varmuza, S. L. & Smiley, J. R. (1985). Signals for site-specific cleavage of HSV DNA: maturation involves two separate cleavage events at sites distal to the recognition sequences. Cell 41, 793802.[Medline]
Wells, R. D., Collier, D. A., Hanvey, J. C., Shimizu, M. & Wohlrab, F. (1988). The chemistry and biology of unusual DNA structures adopted by oligopurine·oligopyrimidine sequences. FASEB J 2, 29392949.
White, C. & Wardropper, A. G. (1997). Genital herpes simplex infection in women. Clin Dermatol 15, 8191.[CrossRef][Medline]
Wohlrab, F., McLean, M. J. & Wells, R. D. (1987). The segment inversion site of herpes simplex virus type 1 adopts a novel DNA structure. J Biol Chem 262, 64076416.
Received 2 September 2002;
accepted 7 November 2002.
HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
INT J SYST EVOL MICROBIOL | MICROBIOLOGY | J GEN VIROL |
J MED MICROBIOL | ALL SGM JOURNALS |