Analysis of Bovine Cytomegalovirus Genome Structure: Cloning and Mapping of the Monomeric Polyrepetitive DNA Unit, and Comparison of European and American Strains

Abstract

SUMMARYThe genome of bovine cytomegalovirus (BCMV) strain 66-P-347 consists of doublestranded, linear DNA with a size of 144 + 6 kb. It contains polyrepetitive DNA (prDNA) segments like five other BCMV strains. The restriction patterns of the prDNA of all six strains are very similar and indicate that monomeric prDNA units are either 1950 bp (class I and Ia), 2350 bp (class II) or 2750 bp (class III) in size. The complete unit sequence of strain 66-P-347 (class II) was cloned in bacteriophage vector M13mp7 and mapped by the restriction enzymes EcoRI, BamHI, BglI, NaeI, SstII and PstI. From these results the restriction maps of the prDNA of the other five strains were deduced. The 400 bp differences in size between the three prDNA classes are a consequence of the appearance of an internal 200 bp sequence being present one-, three-or fivefold in head-to-tail formation. Hybridization of 35S_labelled recombinant phage DNA to Southern blots with DNA of the different strains leads to the conclusion that prDNA units are present as multimers in tandem formation at both genomic termini in the same orientation. The number of terminal repeat units varies between individual molecules of a strain, but the actual terminal sequences are identical.

A multipotential β-1,6-N-acetylglucosaminyl-transferase is encoded by bovine herpesvirus type 4

“…Gammaherpesviruses are lymphotropic viruses represented by Epstein-Barr virus (EBV) and Kaposi's sarcoma-associated herpesvirus (HHV-8). The genome of BHV-4 is a double-stranded 145-kb DNA molecule consisting of a 110-kb central unique coding sequence, called L-DNA, flanked by two stretches of tandem repeats (24,25). These stretches are designated polyrepetitive DNA (prDNA).…”

Structure and function of the prDNA and the genomic termini of the gamma2-herpesvirus bovine herpesvirus type 4.

“…BHV-4 strain 66-p-347 was propagated in Georgia bovine kidney (GBK) cells and BHV-4 particles were prepared from the supernatant of infected cells. Virus DNA isolation has been described previously (Ehlers et al, 1985).…”

“…These stretches have been designated polyrepetitive DNA (prDNA). Using a prDNA unit as a molecular probe, we have (i) identified the genomic termini of BHV-4 as well as restriction fragments representing the junctions between prDNA and unique DNA and (ii) demonstrated that the frequencies of prDNA units at both genomic termini vary between individual molecules of a strain (Ehlers et al, 1985). For the investigated BHV-4 strain 66-p-347 (Storz, Author for correspondence : Michael Goltz.…”

“…Its linear, double-stranded DNA of 144p6 kbp consists of a long central unique part (" 110 kbp) which is flanked by two stretches of tandem repeats with high GjC content (Ehlers et al, 1985) (Fig. 1).…”

DNA-binding Proteins Induced by the Cottontail Rabbit Herpesvirus CTHV

“…Amid this variety, one structural pattern is especially remarkable in that it embodies the extremes of base composition and sequence repetition currently recognized for the herpesvirus group as a whole. Originally discovered in herpesvirus saimiri (Bornkamm et al, 1976), similar genome structures have since been found in other New World primate herpesviruses (reviewed by Fleckenstein & Desrosiers, 1982), several bovine herpesviruses (Ehlers et al, 1985), and two independently isolated cottontail rabbit herpesviruses, herpesvirus sylvilagus (Medveczky et al, 1984;see, however, Rouhandeh & Cohrs, 1987) and CTHV (J. Cebrian et al, personal communication).…”