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Abstract
Interplay between the host and human cytomegalovirus (HCMV) has a pivotal role in the outcome of infection. A region (referred to as UL/b') present in the Toledo strain of HCMV and low passage clinical isolates contains 19 additional genes, which are absent in the highly passaged laboratory strain AD169. Products of the UL/b' genes may determine the manifestations of HCMV infection in vivo. However, little is known about the host factors, which interact with UL/b' proteins. This study was conducted to investigate the function of the HCMV UL136 protein. By yeast two-hybrid screening, the β1 subunit of the host Na+/K+-ATPase (ATP1B1) was identified to be a candidate protein, which interacts with the HCMV UL136 protein. The interaction was further evaluated both in vitro by pull-down assay and in vivo by immunofluorescent co-localization. The results showed that the UL136 protein can interact with ATP1B1 in vitro. Co-localization of UL136-EGFP and ATP1B1-DsRed in cell membranes suggests that ATP1B1 was a partner of the UL136 protein. It can be proposed that the HCMV UL136 protein may have important roles in processes such as cell-to-cell spread, and in maintaining cell osmotic pressure and intracellular ion homeostasis during HCMV infection.
Highlights
Human cytomegalovirus (HCMV), a member of the Betaherpesvirinae subfamily, is widely distributed in human populations
In an immunofluorescent co-localization test, pDsRed/ ATP1B1 was found in the cell membrane, and the pEGFP/ UL136 fusion protein was localized in both the cell membrane and the cytoplasm
Genes in the HCMV UL/b’ region encode several important proteins including the cell-tropic factors (UL131A -128), virion structure (UL132) [2,7], viral latency determinant (UL138), and host cell machinery modulation factors (UL141, 142, 144, and 148) [8,9,10,12,13]. This region is one of the most important targets to elucidate the mechanism of pathogenesis related to clinical HCMV isolates
Summary
Human cytomegalovirus (HCMV), a member of the Betaherpesvirinae subfamily, is widely distributed in human populations. Since 1996, it has been recognized that a 15-kb unique region (UL/b’) contains at least 19 open reading frames (ORFs) These genes are found in clinical HCMV isolates but not in the extensively passaged laboratory AD169 strains, which are dispensable for growth in vitro [5]. The retention of these ORFs in clinical HCMV isolates, and the fact that the virus can replicate in SCID mice implanted with human tissues suggest that the UL/b’ region is required for viral infection in vivo [6]. The UL144 gene encodes a structural homologue of the herpes virus entry mediator, upregulating
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