Identification of an encystation-specific transcription factor, Myb protein in Giardia lamblia

Abstract

The life cycle of Giardia lamblia contains two differentiation processes, encystation and excystation. We performed an experiment to identify the genes induced during encystation using the differential display reverse transcriptase–polymerase chain reaction. Three of twelve isolated cDNA clones that showed increased transcription during encystation were identified to be of the myb2, which encodes a well-known transcriptional factor involved in cellular development and differentiation. The amino acid sequences of the Myb2 protein deduced from the isolated gene revealed that this Myb2 has a DNA binding domain comprising two imperfect repeats at its carboxyl-terminus. The nuclear localization of Myb2 protein during encystation was observed in vivo by expressing a Myb2-GFP fusion protein. In a random site selection experiment, the oligonucleotides bound by rMyb2 contained a conserved sequence of GTTT(G/T)(G/T). Two promoters of the encystation-induced genes, myb2, and cwp1, were also found to bind to rMyb2, whereas gap1, one of the constitutive genes did not.