Functional characterization of all‐trans retinoic acid‐induced differentiation factor (ATRAID)

Abstract

All‐trans retinoic acid‐induced differentiation (ATRAID) factor was first identified in HL60 cells. Several mRNA isoforms exist, but the respective proteins have not been fully characterized. In transfected cells expressing Myc‐Flag‐tagged ATRAID Isoform (Iso) A, B, and C, Iso C was found to be expressed at high levels, Iso A was found to be expressed at low levels due to rapid degradation, and the predicted protein expressed from Iso B was not detected. Iso C was present mainly in an N‐glycosylated form. In subcellular fractionation experiments, Iso C localized to the membranous and nuclear fractions, while immunofluorescence analysis revealed that Iso C is located close to the plasma membrane, mainly in cytoplasmic vesicles and in the Golgi area. We confirm that Iso C colocalizes to some extent with endosomal/lysosomal markers LAMP1 and LAMP2. Furthermore, we show that ATRAID co‐localizes with RAB11, a GTPase associated with recycling endosomes and implicated in regulating vesicular trafficking.

Keywords

• ATRAID
• endosomes
• Golgi
• isoform
• N‐glycosylation
• RAB11