Productive HIV-1 replication is mainly controlled at the transcriptional level by HIV-1 Tat and at the post-transcriptional level by HIV-1 Rev. A number of host factors have been identified to be involved in these processes. Src-associated protein of 68 kDa in mitosis (Sam68) is a multi-functional RNA-binding protein and has been implicated in several important cellular processes. More evidence has accumulated to support an important regulatory function of Sam68 at two distinct steps of HIV-1 gene expression. Sam68 interacts with HIV-1 Rev protein and directly participates in nuclear exportation of HIV-1 unspliced or singly spliced RNA. In addition, Sam68 functions in the cytoplasmic processes of HIV-1 replication such as the translational regulation of HIV-1 RNA. Elucidation of the precise molecular function of Sam68 in HIV-1 gene expression is clearly warranted and is expected to unambiguously establish Sam68 as an important host factor for HIV-1 replication.