Allosteric HIV-1 Integrase Inhibitors Lead to Premature Degradation of the Viral 1 RNA Genome and Integrase in Target Cells 2 3 Running head : Fates of Eccentric HIV-1 Particles in Target Cells 4 5

24 Recent evidence indicates that inhibition of HIV-1 integrase (IN) binding to the 25 viral RNA genome by allosteric integrase inhibitors (ALLINIs) or through mutations within 26 IN yields aberrant particles, in which the viral ribonucleoprotein complexes (vRNPs) are 27 eccentrically localized outside the capsid lattice. These particles are non-infectious and 28 blocked at an early reverse transcription stage in target cells. However, the basis of this 29 reverse transcription defect is unknown. Here we show that the viral RNA genome and 30 IN from ALLINI-treated virions are prematurely degraded in target cells, whereas reverse 31 transcriptase remains active and stably associated with the capsid lattice. The aberrantly 32 shaped cores in ALLINI-treated particles can efficiently saturate and be degraded by a 33 restricting TRIM5 protein, indicating that they are still composed of capsid proteins 34 arranged in a hexagonal lattice. Notably, fates of viral core components follow a similar 35 pattern in cells infected with eccentric particles generated by mutations within IN that 36 inhibit its binding to the viral RNA genome. We propose that IN-RNA interactions allow 37 for packaging of both the viral RNA genome and IN within the protective capsid lattice to 38 ensure subsequent reverse transcription and productive infection in target cells. 39 Conversely, disruption of these interactions by ALLINIs or mutations in IN leads to 40 premature degradation of both the viral RNA genome and IN, as well as the spatial 41 separation of reverse transcriptase from the viral genome during early steps of infection. 42 43