Novel virus-associated proteins encoded by UL112-113 of human cytomegalovirus.
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.
Wang
Shang-Kwei
SK
Department of Microbiology, Institute of Medicine, College of Medicine, Kaohsiung Medical University, 100 Shih-Chuan 1st Road, Kaohsiung 80708, Taiwan, ROC.
Hu
Cheng-Hui
CH
Department of Microbiology, Institute of Medicine, College of Medicine, Kaohsiung Medical University, 100 Shih-Chuan 1st Road, Kaohsiung 80708, Taiwan, ROC.
Lu
Miao-Chan
MC
Department of Microbiology, Institute of Medicine, College of Medicine, Kaohsiung Medical University, 100 Shih-Chuan 1st Road, Kaohsiung 80708, Taiwan, ROC.
Duh
Chang-Yih
CY
Asia-Pacific Ocean Research Center, National Sun Yat-sen University, 70 Lien-Hai Road, Kaohsiung 80424, Taiwan, ROC.
Liao
Pao-Chi
PC
Department of Environmental and Occupational Health, College of Medicine, National Cheng Kung University, 138 Sheng-Li Road, Tainan 704, Taiwan, ROC.
Tyan
Yu-Chang
YC
Department of Environmental and Occupational Health, College of Medicine, National Cheng Kung University, 138 Sheng-Li Road, Tainan 704, Taiwan, ROC.
D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D003587:Cytomegalovirus / Q000235:genetics* / Q000378:metabolism* / Q000502:physiology; D004261:DNA Replication; D015723:Gene Library; D006801:Humans; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D010750:Phosphoproteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D014764:Viral Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D014771:Virion / Q000378:metabolism
D010750:Phosphoproteins; D014764:Viral Proteins