Two-hybrid analysis of the interaction between the UL52 and UL8 subunits of the herpes simplex virus type 1 helicase–primase

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Abstract
Herpes simplex virus type 1 expresses a heterotrimeric helicase–primase, the subunits of which are encoded by the viralUL5,UL8andUL52genes. The interactions of the UL52 protein with the UL8 and UL5 proteins were analysed by using the yeast two-hybrid system. The UL52–UL5 interaction gave a specific but weak signal in the two-hybrid system. In contrast, the UL52–UL8 interaction gave a strong signal in the two-hybrid system. Deletion analysis showed that a 548 amino acid fragment of UL52 (amino acids 366–914) retains the ability to interact with UL8 and that the N-terminal 349 amino acids are dispensable for the interaction. A fragment library screen and co-immunoprecipitation experiments confirmed the deletion analysis results.