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Human herpesvirus-6 rep/U94 gene product has single-stranded DNA-binding activity
Panadda Dhepakson1, Yasuko Mori1, Yun Bao Jiang1
1Department of Microbiology, Osaka University Medical School, 2-2 Yamada-oka, Suita, Osaka 565-0871, Japan1.
The Journal of General Virology
|March 22, 2002
Summary
Researchers characterized the human herpesvirus-6B (HHV-6B) REP protein, finding it localizes to the nucleus and binds single-stranded DNA. This REP protein is expressed at low levels in infected cells.
Area of Science:
- Virology
- Molecular Biology
Background:
- Human herpesvirus-6B (HHV-6B) is a significant human pathogen.
- The HHV-6B rep/U94 gene is homologous to the adeno-associated virus type 2 rep gene, suggesting a potential role in viral replication.
Purpose of the Study:
- To characterize the HHV-6B REP protein.
- To investigate the expression and localization of HHV-6B REP during infection.
- To determine the DNA-binding properties of HHV-6B REP.
Main Methods:
- Production of a monoclonal antibody against HHV-6B REP (anti-REP mAb).
- Immunofluorescence staining in HHV-6-infected MT4 cells.
- Western blot analysis of infected cell lysates.
- Single-stranded DNA binding assays.
Main Results:
- The anti-REP mAb recognized a 56-kDa polypeptide in HHV-6B-infected cells.
- Immunofluorescence showed REP localized to the nucleus, detected from 24 h post-infection and accumulating by 72 h p.i.
- REP expression levels were low, detected in a small percentage of infected cells even when late proteins like glycoprotein H were abundant.
- The REP protein demonstrated binding to single-stranded DNA.
Conclusions:
- The HHV-6B REP protein is expressed in the nucleus of infected cells.
- REP is a low-abundance protein during HHV-6B infection.
- HHV-6B REP possesses DNA-binding capabilities, suggesting a role in viral replication or genome maintenance.