DNA-binding protein associated with herpes simplex virus DNA polymerase

Area of Science:

• Virology
• Molecular Biology
• Immunology

Background:

• Purified herpes simplex virus type 2 (HSV-2) DNA polymerase preparations consistently include two distinct polypeptides.
• The major polypeptide (~150,000 MW) is recognized for its polymerase activity.
• A second polypeptide (~54,000 MW), designated ICSP 34, 35, has been identified in these preparations.

Purpose of the Study:

• To purify the ~54,000 MW polypeptide (ICSP 34, 35) from HSV-2 DNA polymerase preparations.
• To generate specific antibodies against the purified ICSP 34, 35.
• To characterize ICSP 34, 35 and elucidate its relationship with the HSV-2 DNA polymerase.

Main Methods:

• Purification of the ~54,000 MW polypeptide.
• Generation of polyclonal rabbit antisera and monoclonal antibodies against purified ICSP 34, 35.
• Immunological characterization and genetic mapping of HSV-2 polypeptides.

Main Results:

• The ~54,000 MW protein (ICSP 34, 35) was successfully purified.
• Specific antisera and monoclonal antibodies were generated against ICSP 34, 35.
• Characterization revealed a distinct genetic map location for ICSP 34, 35 compared to the DNA polymerase.
• A close association between the two polypeptides was demonstrated.

Conclusions:

• The ~54,000 MW polypeptide (ICSP 34, 35) is a distinct viral protein associated with HSV-2 DNA polymerase.
• Despite separate genetic origins, ICSP 34, 35 and the DNA polymerase exhibit a close functional or physical association.
• Further research is warranted to define the precise role of ICSP 34, 35 in viral DNA replication.