Two-dimensional gel analysis of HSV type 1-induced polypeptides and glycoprotein processing

Area of Science:

• Virology
• Molecular Biology
• Cell Biology

Background:

• Herpes simplex virus type 1 (HSV-1) is a significant human pathogen.
• Understanding HSV-1 protein synthesis and modification is crucial for antiviral development.

Purpose of the Study:

• To comprehensively characterize HSV-1-induced polypeptides in infected BHK cells.
• To investigate post-translational modifications and glycoprotein synthesis pathways.

Main Methods:

• Two-dimensional gel electrophoresis for polypeptide separation and characterization.
• Isoelectric focusing and SDS-polyacrylamide gel electrophoresis for mobility and molecular weight determination.
• Pulse-chase experiments and neuraminidase treatment to study glycoprotein synthesis and modification.

Main Results:

• 230 distinct virus-induced polypeptides were identified in HSV-1 infected BHK cells.
• Several polypeptides, including those in the Vmw43 band (thymidine kinase), were resolved into multiple spots.
• Post-translational modifications were observed, affecting polypeptide mobility.
• Glycoproteins gB, gC, and gD were found to contain sialic acid.
• Synthesis of gB and gD involves at least 15 and 10 discrete steps, respectively.

Conclusions:

• HSV-1 infection induces a complex array of polypeptides with evidence of significant post-translational modification.
• The study provides detailed insights into the synthesis and modification of key HSV-1 glycoproteins, including sialic acid incorporation.