Perturbing HIV-1 genomic RNA subcellular localization inhibits virus particle production

14 15 16. CC-BY-NC-ND 4.0 International license peer-reviewed) is the author/funder. It is made available under a The copyright holder for this preprint (which was not. Abstract. mRNA subcellular localization is a crucial determinant of eukaryotic gene 17 expression. For retroviruses including HIV-1, the unspliced genomic RNA (gRNA) 18 serves both as the mRNA encoding Gag/Gag-Pol capsid proteins and the genetic 19 material packaged by Gag into virions assembling at the plasma membrane. Gag is 20 sufficient to drive the assembly of virus-like particles in the absence of gRNA binding. 21 Thus, what role gRNA cytoplasmic trafficking plays during assembly is unknown. Here 22 we demonstrate that aberrantly tethering HIV-1 gRNAs to membranes or the actin 23 cytoskeleton alters Gag subcellular distribution and reduces virus particle production. 24 This block was restricted to gRNAs competent for Gag synthesis and mapped to the 25 Rev response element; a cis-acting RNA structure that regulates gRNA nuclear export. 26 These results expose an unexpected mechanistic link between HIV-1 gRNA nuclear 27 history, cytoplasmic trafficking, and Gag assembly. Perturbing viral mRNA subcellular 28 distribution could represent a novel antiviral strategy. 29 30 31. CC-BY-NC-ND 4.0 International license peer-reviewed) is the author/funder. It is made available under a The copyright holder for this preprint (which was not .