Structural characterization of the Molluscum contagiosum virus genome

Parr, Ronald P.; Burnett, Joseph W.; Garon, Claude F.

doi:10.1016/0042-6822(77)90141-6

Cited by 52 publications

(20 citation statements)

References 16 publications

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“…However, it differs in its susceptibility to cleavage by endonuclease Sinai (recognition sequence CCCGGG) which generates more than 25 cuts in MCV DNA, in contrast to the few in Orthopoxvirus DNA (Mackett & Archard, 1979). This result would be expected from the higher G + C content of MCV DNA (estimated as 60~o) compared to Orthopoxvirus DNA (Parr et al, 1977).…”

Section: Discussionmentioning

confidence: 64%

“…(1986) was from the only genital lesion in their series. Parr et al (1977) examined virus from a total of 10 lesions, two of which were genital; in both these cases the BamHI cleavage patterns of virus DNA correspond to MCV II, although they found this virus in other sites also. In contrast, we observed a total of six instances of MCV II, only one of which was of genital origin; MCV I was isolated from four other genital lesions.…”

Section: Discussionmentioning

confidence: 99%

“…Attempts to propagate virus in vitro have not been successful (Postlethwaite, 1970). Parr et al (1977) studied the MCV genome by electron microscopy and showed by partial denaturation that it is a terminally cross-linked molecule of about 180 kb. They also demonstrated three different BamHI cleavage patterns of DNA from MCV isolates.…”

Section: Introductionmentioning

confidence: 99%

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Characterization and Physical Mapping of Molluscum Contagiosum Virus DNA and Location of a Sequence Capable of Encoding a Conserved Domain of Epidermal Growth Factor

Porter

Archard

1987

Journal of General Virology

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SUMMARYDNA from Molluscum contagiosum virus (MCV) isolates was analysed by restriction endonuclease cleavage, revealing two virus subtypes. Physical maps of cleavage sites for BamHI, Clal and HindIII were constructed, and found to differ extensively between the two subtypes. MCV DNA was similar to Orthopoxvirus DNA with respect to size, terminal cross-linking and the presence of inverted terminal repetitions, but did not hybridize with vaccinia virus DNA. The genomes of the two MCV subtypes crosshybridized and were colinear except for two small regions. There was sequence homology between DNA from corresponding map regions of the MCV subtypes but, in contrast to Orthopoxvirus DNA, no conservation of restriction sites. A synthetic oligonucleotide probe representing a conserved domain of epidermal growth factor, c~-transforming growth factor and the vaccinia growth factor identified equivalent regions of both MCV genomes as having the potential to encode this domain. This locus is similar to the position of the vaccinia growth factor gene in vaccinia virus DNA. Thus MCV may induce epidermal cell proliferation and tumourigenesis by expression of an epidermal growth factor-like polypeptide. INTRODUCTIONMolluscum contagiosum virus (MCV) is an unclassified poxvirus of man which induces benign skin tumours. Transmission is mechanical and, in adults, may be venereal. The skin lesion consists of a localized mass of hypertrophied and hyperplastic epidermis extending down into the underlying dermis and projecting above the surface as a papule. The rate of cell division in the basal layer is several times that of normal skin. The lower cells of the stratum spinosum contain characteristic cytoplasmic inclusion bodies resulting from virus replication, which enlarge as the infected cells migrate through the stratum granulosum towards the surface. These molluscum bodies are trapped in the horny layer by a fibrous network which dissolves in the centre of the lesion forming a central core composed primarily of virus. The disease is selflimiting but lesions may be curetted or the virus-rich core collected. Attempts to propagate virus in vitro have not been successful (Postlethwaite, 1970). Parr et al. (1977) studied the MCV genome by electron microscopy and showed by partial denaturation that it is a terminally cross-linked molecule of about 180 kb. They also demonstrated three different BamHI cleavage patterns of DNA from MCV isolates. Dural et aL (1986) have reported two viral subtypes on the basis of different cleavage patterns.MCV is one of several tumourigenic poxviruses. Others are Shope rabbit fibroma virus and Yaba monkey tumour virus which induce fibromas and histiocytomas respectively. Induction of epidermal cell proliferation by poxviruses is of interest following recent studies demonstrating that a vaccinia virus Mr 19 000 (19K) polypeptide is structurally and functionally homologous to epidermal growth factor (EGF) and to c~-transforming growth factor (~-TGF) (Brown et al.,

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Section: Discussionmentioning

confidence: 64%

Section: Discussionmentioning

confidence: 99%

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Characterization and Physical Mapping of Molluscum Contagiosum Virus DNA and Location of a Sequence Capable of Encoding a Conserved Domain of Epidermal Growth Factor

Porter

Archard

1987

Journal of General Virology

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“…Between 5-20% of HIV patients have MC infection. The incubation period for MC has been reported to be between 14 and 50 days, although there are reports of newborns having lesions as early as 7 days postpartum (6).An individual infected with Molluscum contagiosum virus can also spread the infection via autoinoculation. The virus infects epidermal keratinocytes and viral replication occurs in the cytoplasm of these cells (7).…”

Section: Introductionmentioning

confidence: 99%

<i>Molluscum contagiosum</i> virus infection amongst plwha in ibadan, Nigeria

Fayemiwo¹,

Adesina²,

Akinyemi³

et al. 2013

Af J Clin Exp Micro

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“…Partial denaturation mapping of vaccinia virus DNA indicated that the base composition at the two ends of the molecule is similar (1,2). Observations that restriction fragments from opposite ends of rabbitpox virus DNA crosshybridize (3) and that similar size restriction fragments are generated from both ends of the vaccinia virus genome (R. Wittek, personal communication) suggested the presence of a terminal repetition in poxvirus DNA.…”

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confidence: 99%

Visualization of an inverted terminal repetition in vaccinia virus DNA.

Garon¹,

Barbosa²,

Moss³

1978

Proc. Natl. Acad. Sci. U.S.A.

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An inverted terminal repetition was observed in DNA molecules extracted from vaccinia virus. The repeated sequence was visualized by (i) nicking the hairpin loops present at the ends of vaccinia virus DNA, (ii) separating the strands of DNA by alkali denaturation, (iii) allowing the single strands to self-anneal, and (iv) examining the DNA with an electron microscope. Single-stranded circular molecules, each of which contained a duplex projection (3.54 i 0.12 M&m) representing the terminal repetition, readily formed. Similar size projections were also seen in heteroduplex structures formed by crosshybridization of the separated strands of the two terminal HindIll restriction fragments. Based on contour length measurements and the electrophoretic mobility of the isolated inverted terminal repetition, a molecular weight of approximately 6.9 X 106, equivalent to about 10,500 nucleotide base pairs, was estimated. Evidence was obtained from DNA*RNA hybridization studies that the terminal repetition is transcribed. The genome of vaccinia virus appears, in the electron microscope, as a typical linear duplex DNA molecule of approximately 122 X 106 daltons (1). Upon complete denaturation, however, single-stranded circles measuring twice the length of the linear duplex are observed (1). The latter finding, together with the failure of the complementary strands to separate upon sedimentation through an alkaline sucrose gradient (1), indicate that the ends of the DNA duplex form a continuous hairpin loop or are crosslinked in some unknown manner. A similar genome structure has been found in molluscum contagiosum virus (2) and may be characteristic of all poxviruses.Partial denaturation mapping of vaccinia virus DNA indicated that the base composition at the two ends of the molecule is similar (1, 2). Observations that restriction fragments from opposite ends of rabbitpox virus DNA crosshybridize (3) and that similar size restriction fragments are generated from both ends of the vaccinia virus genome (R. Wittek, personal communication) suggested the presence of a terminal repetition in poxvirus DNA. By using a single-strand-specific endonuclease (4) to cleave the hairpin loops at the two ends of the vaccinia virus genome (1) and then self-annealing the separated strands of DNA, we now provide direct visual evidence for an inverted terminal repetition approximately 10,500 nucleotide base pairs long. Evidence that at least a portion of the repeated sequence is transcribed will also be presented.MATERIALS AND METHODS Preparation of Vaccinia Virus DNA. HeLa S-3 cells were infected with cloned vaccinia virus (strain WR) and virions were purified without sonication as described (5). DNA was extracted as described (6) except that digestion with 1 mg of proteinase K (EM Laboratories Inc., Elmsford, NY) per ml for 5 hr was followed by repeated phenol extraction and dialysisThe publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in Lab...

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Structural characterization of the Molluscum contagiosum virus genome

Cited by 52 publications

References 16 publications

Characterization and Physical Mapping of Molluscum Contagiosum Virus DNA and Location of a Sequence Capable of Encoding a Conserved Domain of Epidermal Growth Factor

Characterization and Physical Mapping of Molluscum Contagiosum Virus DNA and Location of a Sequence Capable of Encoding a Conserved Domain of Epidermal Growth Factor

<i>Molluscum contagiosum</i> virus infection amongst plwha in ibadan, Nigeria

Visualization of an inverted terminal repetition in vaccinia virus DNA.

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