Human Cytomegalovirus DNA Sequences with Homologies to the Cellular Genome

Abstract

SUMMARYDNA from a cosmid-cloned gene library of human cytomegalovirus (HCMV) strain AD169 was found to share sequence homologies with cellular DNA of various origins. Viral DNA fragments subcloned in plasmid vectors enabled localization of the homologous sequences to five regions of the HCMV genome. Four of these regions were found in the long unique (UL) segment of the viral genome (EcoRI fragments R, I and b, and HindlII fragment S); one region with virus-cell homology was located in the terminal inverted repeats (EcoRI fragments O and H). All hybridization reactions were carried out under stringent reannealing conditions (18 °C to 21-5 °C below average T m of HCMV DNA). Fragments from these five HCMV DNA regions did not crosshybridize with each other. The sequence homologies were seen in DNA from normal human placental tissue, liver autopsy material, intestinal tissue, white blood cells, and colon carcinoma cells, and in a series of haematopoietic cell lines. Sequence homologies were also detected in DNA from owl monkeys and Chinese hamsters, but not in mouse DNA. They were not related to human Alu repeats or cloned human c-myc sequences.

Primer-mediated enzymatic amplification of cytomegalovirus (CMV) DNA. Application to the early diagnosis of CMV infection in marrow transplant recipients.

“…For studies of CMV excretion, 12 ml of urine or saliva (collected in saline) were centrifuged for 1 h at 38,000 rpm (180,000 g) in an SW-41 rotor in a centrifuge (Beckman Instruments, Inc., Palo Alto, CA). The concentrated viral pellets were then lysed and DNA was extracted (22 (24,25) and it has no reported homology with human DNA (26,27). Synthesis of the primers and probe were performed on a DNA synthesizer (model 380 A; Applied Biosystems, Foster City, CA) using published sequence data (24) and the methoxyphosphoramidite method (28).…”

The status of CMV as a human pathogen

“…Since regions of CMV DNA are now known to bind avidly to human DNA (Peden, Mounts & Hayward, 1982;Ruger, Bornkamm & Fleckenstein, 1984), the results claiming to detect CMV DNA or RNA must be assumed to be unreliable until the experiments have been repeated using cloned sequences of the virus genome which lack cross-reactivity with host DNA. Likewise, the studies of virus-specified proteins within KS tissue must be repeated using monoclonal antibodies of defined specificity before the results can be accepted.…”

Human Cytomegalovirus: Recent Aspects from Molecular Biology

“…ROger et al (1984a) and Shaw et al (1985) found five regions with cell-related nucleotide sequences in the genome of HCMV strain AD 169. It has been determined that the nucleotide sequence homologies of HCMV DNA are different from those in simian cytomegalovirus (SCMV) strains, where tandem repeats of CA dinucleotides appeared to hybridize with highly repetitive sequence elements in cellular DNA (Jeang & Hayward, 1983).…”