HIV-1 capsids mimic a microtubule regulator to coordinate early stages of infection

While the microtubule end-binding protein,EB1 facilitates early stages ofHIV-1 infection, how it does so remains unclear. Here, we show that beyond its effects on microtubule acetylation,EB1 also indirectly contributes to infection by delivering the plus-end tracking protein (+TIP), cytoplasmic linker protein 170 (CLIP170) to the cell periphery.CLIP170 bound to intactHIV-1 cores orin vitroassembled capsid-nucleocapsid complexes, whileEB1 did not. Moreover, unlikeEB1 and several other +TIPs,CLIP170 enhanced infection independently of effects on microtubule acetylation. Capsid mutants and imaging revealed thatCLIP170 boundHIV-1 cores in a manner distinct from currently known capsid cofactors, influenced by pentamer composition or curvature. Structural analyses revealed anEB-like +TIP-binding motif within the capsid major homology region (MHR) that binds SxIPmotifs found in several +TIPs, and variability across thisMHRsequence correlated with the extent to which different retroviruses engageCLIP170 to facilitate infection. Our findings provide mechanistic insights into the complex roles of +TIPs in mediating early stages of retroviral infection, and reveal divergent capsid-basedEB1 mimicry across retroviral species.