Recent genome-wide screens have highlighted an important role for transportin-3 in human immunodeficiency virus type 1 (HIV-1) infection and preintegration complex (PIC) nuclear import. HIV-1 integrase moreover interacted with recombinant transportin-3 protein under conditions whereby Moloney murine leukemia virus (MLV) integrase failed to do so, suggesting that integrase-transportin-3 interactions might underscore active retroviral PIC nuclear import. Here we correlate infectivity defects in transportin-3 knockdown cells with in vitro protein binding affinities for an expanded set of retroviruses that include simian immunodeficiency virus (SIV), bovine immunodeficiency virus (BIV), equine infectious anemia virus (EIAV), feline immunodeficiency virus (FIV), and Rous sarcoma virus (RSV) to critically address the role of integrase-transportin-3 interactions in viral infection. Lentiviruses with the exception of FIV display a requirement for transportin-3 in comparison to MLV and RSV, yielding an infection-based dependency ranking of SIV > HIV-1 > (BIV, EIAV) > (MLV, RSV, FIV). In vitro pulldown and surface plasmon resonance assays by contrast define a notably different integrase-transportin-3 binding hierarchy: (FIV, HIV-1, BIV) > (SIV, MLV) > EIAV. Our results therefore fail to support a critical role for integrase binding in dictating transportin-3 dependency during retrovirus infection. In addition to integrase, capsid has been highlighted as a retroviral nuclear import determinant. Accordingly, MLV/HIV-1 chimera viruses pinpoint the genetic determinant of sensitization to transportin-3 knockdown to the HIV-1 capsid protein. We therefore conclude that capsid, not integrase, is the dominant viral factor that dictates transportin-3 dependency during HIV-1 infection.