Characterization of human cytomegalovirus UL145 and UL136 genes in low-passage clinical isolates from infected Chinese infants

Abstract

SummaryBackgroundHuman cytomegalovirus (HCMV) is a leading cause of morbidity and mortality in immunocompromised individuals. The unique long b’ (ULB’) region of HCMV contains at least 19 open reading frames (ORFs); however, little is known about the function of UL145 and UL136. We characterized UL145 and UL136 in low-passage clinical isolates from Chinese infants.Material/MethodsThe clinical strains of HCMV were recovered from the urine from HCMV-infected infants. Human embryonic lung fibroblasts (HELFs) were infected with clinical isolates of HCMV, and the viral DNA and mRNA for UL145 and UL136 were analyzed by polymerase chain reaction (PCR) and sequencing techniques. We also predicted the structure and function of UL145 and UL136 proteins.ResultsSixty-two Chinese infants infected with HCMV were recruited into this study and the clinical isolates were recovered from the urine. Two strains among the low-passage isolates, D2 and D3, were obtained. The UL145 and UL136 sequences were deposited with GenBank under accession numbers of DQ180367, DQ180381, DQ180377, and DQ180389. The mRNA expression of both UL145 and UL136 was confirmed by reverse transcription (RT-PCR) assays. UL145 was predicted to contain 1 protein kinase C phosphorylation site, 2 casein kinase II phosphorylation sites and a zinc finger structure. UL136 was predicted to contain a protein kinase C phosphorylation site, N-myristoylation site, cAMP- and cGMP-dependent protein kinase phosphorylation site and tyrosine kinase II phosphorylation site. Both UL145 and UL136 are highly conserved.ConclusionsUL145 may act as an intranuclear regulating factor by direct binding to DNA, while UL136 may be a membrane receptor involving signal transduction.

Disease Activity-Associated Alteration of mRNA m5 C Methylation in CD4+ T Cells of Systemic Lupus Erythematosus

“…Recently, m 5 C methylation in eukaryotic cells has been identified to largely depend on the function of the known "writer" NSUN2, which functions as an mRNA methyltransferase (Bohnsack et al, 2019;Yang Y. et al, 2019). Moreover, it was reported that NSUN2 affects cell biological processes including cell proliferation, migration, and mRNA metabolism by regulating mRNA m 5 C modification (Wang et al, 2011;Courtney et al, 2019). Compared to that in HCs, the NSUN2 expression (including mRNA and protein) in CD4 + T cells of patients with SLE was significantly lower, which further confirmed the Bis-Seq data of overall m 5 C levels in SLE.…”

Cellular Cullin RING Ubiquitin Ligases: Druggable Host Dependency Factors of Cytomegaloviruses

“…The most recently described member of HCMV-encoded DDB1/CRL-interacting proteins is pUL145. In contrast to the adjacent genes UL144 and UL146 , UL145 is highly conserved among different HCMV strains, suggesting its relevance for viral replication as well as the conservation of its interaction partner(s) [120,139,140,141]. The UL145 gene gives rise to monocistronic as well as polycistronic transcripts [141,142].…”

“…The prediction of motifs for posttranslational modifications suggested a protein kinase C phosphorylation motif and one or two casein kinase II phosphorylation site(s) [139,141]. Additionally, Wang et al proposed the existence of a zinc finger structure [140]. However, to our knowledge, a crystal structure is not yet available.…”

Detection of Cytomegalovirus (CMV) Infection in Wheezing Infants by Urine DNA and Serum IgG Testing

“…Mononuclear cells were isolated from the blood samples using TFicoll separation solution (Shanghai Hua Jing Biotech Company). The urine samples were centrifuged for 10 minutes at 1,500 rpm and then the sediment was collected [ 17 ]. The DNA from mononuclear cells and from the sediment of urine samples were extracted using a DNA extraction kit according to the manufacturer’s instructions.…”