The human cytomegalovirus UL78 gene is highly conserved among clinical isolates, but is dispensable for replication in fibroblasts and a renal artery organ-culture system.
The human cytomegalovirus (HCMV) UL78 ORF is considered to encode a seven-transmembrane receptor. However, neither the gene nor the UL78 protein has been characterized so far. The objective of this study was to investigate the UL78 gene and to clarify whether it is essential for replication. UL78 transcription was activated early after infection, was inhibited by cycloheximide but not by phosphonoacetic acid, and resulted in a 1.7 kb mRNA. Later in the replication cycle, a second mRNA of 4 kb evolved, comprising the UL77 and UL78 ORFs. The 5' end of the UL78 mRNA initiated 48 bp upstream of the translation start and the polyadenylated tail started 268 bp downstream of the UL78 translation stop codon within the UL79 ORF. By using bacterial artificial chromosome technology, a recombinant HCMV lacking most of the UL78 coding region was constructed. Successful reconstitution of the UL78-deficient virus proved that the gene was not essential for virus replication in fibroblasts. The deletion also did not reduce virus replication in ex vivo-cultured sections of human renal arteries. Analysis of viral proteins at different stages of the replication cycle confirmed these results. Among clinical HCMV isolates, the predicted UL78 protein was highly conserved. However, an accumulation of different single mutations could be found in the N-terminal region and at the very end of the C terminus. Due to the absence of an in vivo HCMV model, the role of UL78 in the pathogenesis of HCMV infection in humans remains unclear.
Michel
Detlef
D
Abteilung Virologie, Universitätsklinikum, Albert-Einstein-Allee 11, 89081 Ulm, Germany.
Milotić
Irena
I
Clinic for Infectious Diseases, Clinical Hospital Centre, Cambierieva 17, 51000 Rijeka, Croatia.
Wagner
Markus
M
Harvard Medical School, NRB 836, Department of Pathology, 77 Avenue Louis Pasteur, Boston, MA 02115, USA.
Vaida
Bianca
B
Abteilung Virologie, Universitätsklinikum, Albert-Einstein-Allee 11, 89081 Ulm, Germany.
Holl
Jens
J
Abteilung Virologie, Universitätsklinikum, Albert-Einstein-Allee 11, 89081 Ulm, Germany.
Ansorge
Ramona
R
Abteilung Virologie, Universitätsklinikum, Albert-Einstein-Allee 11, 89081 Ulm, Germany.
Mertens
Thomas
T
Abteilung Virologie, Universitätsklinikum, Albert-Einstein-Allee 11, 89081 Ulm, Germany.
D000328:Adult; D000368:Aged; D002478:Cells, Cultured; D002648:Child; D003587:Cytomegalovirus / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D005347:Fibroblasts; D005814:Genes, Viral / Q000502:physiology*; D006801:Humans; D007223:Infant; D008969:Molecular Sequence Data; D008970:Molecular Weight; D016366:Open Reading Frames; D009924:Organ Culture Techniques; D026723:Polyadenylation; D017433:Protein Structure, Secondary; D012333:RNA, Messenger / Q000096:biosynthesis / Q000737:chemistry; D012077:Renal Artery; D014779:Virus Replication / Q000235:genetics*
D012333:RNA, Messenger