Interaction Cloning and Characterization of the cDNA Encoding the Human Prenylated Rab Acceptor (PRA1)

• Published in: Biochemical and biophysical research communications 1999-05, Vol.258 (3), p.657-662
• Main Authors: Bucci, Cecilia, Chiariello, Mario, Lattero, Daniela, Maiorano, Monica, Bruni, Carmelo B.
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Animals; Base Sequence; Carrier Proteins - genetics; Carrier Proteins - metabolism; Cell Line; Cloning, Molecular; Cricetinae; DNA, Complementary; GTP-Binding Proteins; Humans; Membrane Proteins; Molecular Sequence Data; Rats; Sequence Homology, Amino Acid; Vesicular Transport Proteins
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1006/bbrc.1999.0651

Rab proteins are small GTPases involved in the regulation of intracellular membrane traffic in mammalian cells. In order to find Rab-interacting proteins we performed a two-hybrid screening using a human brain cDNA library. Here we report the isolation of a full-length human cDNA clone coding for a protein of 185 amino acids. This protein interacts strongly with the Rab4b, Rab5a, and Rab5c proteins and weakly with Rab4a, Rab6, Rab7, Rab17, and Rab22 in the two-hybrid assay. Comparison with the Data Bank revealed that this clone represents the human homolog of the previously isolated rat Prenylated Rab Acceptor (rPRA1). Analysis of mRNA expression shows a single abundant mRNA of about 0.8 kb ubiquitously expressed. Western blot analysis of the overexpressed protein shows a band of the expected size equally distributed between cytosol and membranes.