pUL37 R2 is essential for sustained retrograde motion during ingress.
2017-12-07T18:59:25Z (GMT) by
<p>Dorsal root ganglion (DRG) sensory neuron explants were infected in 2 ml of media with 3.5 x 10<sup>7</sup> PFU/ml of PRV (WT and R2 mutant) or 1.3 x 10<sup>7</sup> PFU/ml of HSV-1 (WT and R2 mutant). Viruses encoded a pUL25/mCherry or pUL25/GFP fusion to provide imaging of individual capsids in living cells. Capsid axonal transport was recorded by time-lapse fluorescence imaging between 3–4 hpi. More than 90 capsids were analyzed per experiment across three biological replicates. <b>(A)</b> Fraction of time that individual wild-type (WT) and R2-mutant (R2) capsids moved retrogradely (white), anterogradely (gray), or were motionless (black). Error bars are s.d. <b>(B)</b> Net displacement of capsids over a period of 10 seconds. Positive values indicate movement towards neuronal soma (retrograde displacement). Error bars are s.d. (****, p < 0.0001 based on two-tailed unpaired <i>t</i> test). <b>(C)</b> Representative kymographs of axonal transport in neurons. Distance and time are represented on the x and y axis respectively. <b>(D)</b> Delivery of capsids to nuclear rims at 3–4 hpi following co-infection of DRGs with a pUL25/GFP tagged wild-type (WT) virus and either WT or R2 mutant virus encoding a pUL25/mCherry capsid tag.</p>