The Nir/rdgB family of proteins has been identified in a variety of eukaryotic organisms, ranging from worms to mammals. The Drosophila retinal degeneration B (rdgB), a protein that is required for photoreceptor cell viability and light response, was the first to be identified. It consists an amino-terminal phosphatidylinositol (PI)-transfer domain and was proposed to play an essential role in photoreceptor membrane renewal and biogenesis. The other Nir/rdgB family members are functionally and structurally related to the Drosophila homolog and are implicated in regulation of lipid trafficking, metabolism, and signaling. Recent advances have revealed that Nir/rdgB proteins are also involved in regulation of cytoskeletal elements. Thus, these family members exert a broad spectrum of cellular functions and are involved in multiple cellular processes. The physiological functions of these closely related proteins are described in this review.