Two-hybrid analysis of the interaction between the UL52 and UL8 subunits of the herpes simplex virus type 1 helicase-primase.
Herpes simplex virus type 1 expresses a heterotrimeric helicase-primase, the subunits of which are encoded by the viral UL5, UL8 and UL52 genes. 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.
Constantin
Nicoleta
N
Department of Biochemistry, University of Arizona, Biological Sciences West Building, 1041 E. Lowell Street, Tucson, AZ 85721-0088, USA1.
Dodson
Mark S
MS
Department of Biochemistry, University of Arizona, Biological Sciences West Building, 1041 E. Lowell Street, Tucson, AZ 85721-0088, USA1.
D000595:Amino Acid Sequence; D000818:Animals; D004265:DNA Helicases / Q000737:chemistry* / Q000235:genetics / Q000502:physiology; D019915:DNA Primase / Q000737:chemistry* / Q000235:genetics / Q000502:physiology; D018259:Herpesvirus 1, Human / Q000201:enzymology*; D008969:Molecular Sequence Data; D011233:Precipitin Tests; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D018411:Spodoptera; D013329:Structure-Activity Relationship
D011993:Recombinant Fusion Proteins; D019915:DNA Primase; D004265:DNA Helicases