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Originally published as JGV in Press, 10.1099/vir.0.013037-0 on August 12, 2009 Originally published as JGV in Press, 10.1099/vir.0.013037-0 on August 5, 2009 J Gen Virol 90 (2009), 2840-2848; DOI 10.1099/vir.0.013037-0

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Novel virus-associated proteins encoded by UL112–113 of human cytomegalovirus

Shang-Kwei Wang1, Cheng-Hui Hu1, Miao-Chan Lu1, Chang-Yih Duh2, Pao-Chi Liao3 and Yu-Chang Tyan3,{dagger}

1 Department of Microbiology, Institute of Medicine, College of Medicine, Kaohsiung Medical University, 100 Shih-Chuan 1st Road, Kaohsiung 80708, Taiwan, ROC.
2 Asia-Pacific Ocean Research Center, National Sun Yat-sen University, 70 Lien-Hai Road, Kaohsiung 80424, Taiwan, ROC
3 Department of Environmental and Occupational Health, College of Medicine, National Cheng Kung University, 138 Sheng-Li Road, Tainan 704, Taiwan, ROC

Correspondence
Shang-Kwei Wang
skwang{at}kmu.edu.tw

Evidence suggests that the products of the human cytomegalovirus (HCMV) UL112–113 genes are involved in viral DNA replication during lytic infection. A polyclonal antibody was raised against the UL112 open reading frame (ORF) to characterize its function in detail. Immunoblots utilizing the UL112 antibody identified seven distinct protein bands (p20, p26, p28, p34, p43, p50 and p84) expressed during the HCMV infectious cycle. After screening a cDNA library constructed from cells 72 h after infection with HCMV, only four different cDNA protein-producing constructs were obtained, and their ORFs corresponded to p34, p43, p50 and p84. The proteins p20, p26 and p28 were further shown to be selectively included within mature HCMV particles, virions, non-infectious enveloped particles and dense bodies. Immunoaffinity protein purification was used to prepare the samples for liquid chromatography coupled to tandem mass spectrometry. This analysis revealed that p20, p26 and p28 were derived from the UL112 ORF, most likely through post-translational proteolytic cleavage.

{dagger}Present address: Department of Medical Imaging and Radiological Sciences, Kaohsiung Medical University, Kaohsiung 80708, Taiwan, ROC.

The GenBank/EMBL/DDBJ accession numbers for the complete nucleotide sequences of the full-length cDNAs for p34, p43, p50 and p84 obtained in this study are U57433 [GenBank] , U57434 [GenBank] , U57431 and U57432 [GenBank] , respectively.

Supplementary methods, results and figures are available with the online version of this paper.






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