Transcriptional regulation and influence on replication of the human cytomegalovirus UL138 1.4 kb transcript.

ABSTRACT

Human cytomegalovirus (HCMV) exists in a latent form in hematopoietic progenitors and undifferentiated cells of myeloid lineage. Protein UL138, encoded by the UL/b' region of the viral genome, serves an important role in the establishment and/or persistence of HCMV latency. However, little information about transcriptional regulation of the UL138 gene has been reported thus far. In the present study, the transcriptional regulation element (TRE) of the 1.4 kb UL138 region was identified using a series of dual­luciferase constructs that contain 5' truncated deletion fragments located upstream of the transcription start site of the gene. The results demonstrated that the region from nucleotide 188995­188962 of the Han strain genome exhibits promoter activity and harbors the functional binding motif for transcription factor AP­1 (Ap­1). Using electrophoretic mobility shift assays the physical interaction of the transcription factor to a minimal essential core sequence was demonstrated. Northern blotting revealed that deletion of the TRE in a HCMV bacterial artificial chromosome or inhibition of Ap­1 using RNA interference eliminated or reduced the production of the UL138 1.4 kb mRNA transcript in infected human embryonic lung fibroblast cells (HELF). Deletion of the UL138 1.4 kb transcript resulted in acceleration of HCMV replication in HELF cells. To the best of the authors' knowledge, the present study is the first to analyze the transcriptional regulation of the UL138 1.4 kb transcript. Knowledge of the transcriptional regulation of the UL138 gene will enhance understanding of its mechanism in HCMV latency.