Fig. 1

HSV1 encodes three tail-anchored proteins that interact with Asna1/TRC40. a Schematic diagrams show the domain organization of pUL34, pUL56 and pUS9. The transmembrane domain (TMD) of pUL34 (residues 252–272), pUL56 (residues 211–231) and pUS9 (residues 69–89) and the hydrophobicity plots generated by TMpred (http://embnet.vital-it.ch/software/TMPRED_form.html) are depicted. b The yeast two-hybrid (Y2H) system was used to analyse the interaction of Asna1 and the HSV1 encoded tail-anchored (TA) proteins pUL34, pUL56 and pUS9. pUL45, a type II membrane protein with an N-terminal transmembrane protein was used as control. Asna1 fused to the Gal4 activation domain (AD) was tested for interaction with pUL34, pUL56, pUS9 and pUL45 fused to the Gal4 DNA-binding domain (DBD). Interaction of proteins is indicated by transcriptional activation of the HIS3 reporter gene enabling growth (black squares) or no growth (grey squares) of yeast cells on selective media. c To determine the subcellular distribution of Asna1/TRC40 during HSV1 infection, HeLa cells were mock treated or infected with HSV1(F) at an MOI of 1 for 12 h followed by IF analysis using Asna1/TRC40- and Calreticulin-specific antibodies followed by secondary reagents. Nuclei were visualized by DAPI staining