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Characteristics of DNA‐Binding Activity of Human Cytomegalovirus ppUL44
Author(s) -
Hwang Eungsoo,
Kim Jinhee,
Jong Hyunsoon,
Park Jaewon,
Park Chunggyu,
Cha Changyong
Publication year - 2000
Publication title -
microbiology and immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.664
H-Index - 70
eISSN - 1348-0421
pISSN - 0385-5600
DOI - 10.1111/j.1348-0421.2000.tb02570.x
Subject(s) - monoclonal antibody , biology , microbiology and biotechnology , human cytomegalovirus , antigen , dna , affinity chromatography , antibody , biochemistry , gene , enzyme , genetics , immunology
Human cytomegalovirus (HCMV)‐specific monoclonal antibody, SCMVM34, recognizes the early antigen encoded by UL44 of HCMV. This antigen is confined to the nucleus of HCMV‐infected cells. This study was performed to characterize the DNA‐binding activity of the protein encoded by UL44 of HCMV. The nuclear and cytoskeletal fraction of HCMV‐infected cells was subjected to 0.4 m NaCl extraction, DEAE‐Sephacel ion exchange chromatography, DNA‐cellulose chromatography and SDS‐PAGE analysis with monitoring of the reactive protein using SCMVM34 monoclonal antibody. The molecular weights of the resultant proteins were found to be 34, 40 and 52 kDa. The internal peptide fragments were isolated by tryptic digestion and reverse‐phase HPLC. The internal amino acid sequence analysis of the peptides from the HPLC profile revealed that the antigen recognized by SCMVM34 monoclonal antibody was ppUL44. The reactive antigen began to be eluted from 250 m m NaCl (Tris‐HCl pH 7.4) in DNA cellulose. The 34 kDa protein seems to bind to DEAE more tightly than the 52 kDa protein. The surface charge of 34 kDa might be more basic. Conclusively, the antigen recognized by SCMVM34 was the protein encoded by HCMV UL44, which was localized in the nuclei after HCMV infection, and was the DNA‐binding protein with the characteristic that the surface charge of the molecule was more basic, as the molecular weights of the protein were decreased.