Marek's disease alphaherpesvirus (MDV) RLORF4 is not required for expression of glycoprotein C and interindividual spread

Marek's disease virus (MDV) is a lymphotropic alphaherpesvirus that causes Marek's disease (MD) in chickens. RLORF4 is a MDV-specific gene that is systematically deleted during attenuation of MDV in vitro. Concomitantly, the expression of glycoprotein C (gC) is diminished during attenuation, suggesting these two changes may be linked. Original studies in which RLORF4 was deleted utilized an infectious clone that lacked gC expression due to a frame-shift mutation within the gC open reading frame (UL44). Here, we utilized an infectious clone in which gC expression was restored to test our hypothesis that RLORF4 is important for expression of MDV gC, and subsequently, interindividual spread. Contrary to our hypothesis, gC expression was unaltered during both in vitro and in vivo replication of RLORF4-null MDV and was able to efficiently transmit from chicken to chicken, conclusively showing that RLORF4 does not regulate gC expression and is not required for horizontal transmission.