Functional Identification of a Nuclear Localization Signal of MYB2 Protein in Giardia lamblia
MYB2 protein was identified as a transcription factor that showed encystation-induced expression in <i>Giardia lamblia</i>. Although nuclear import is essential for the functioning of a transcription factor, an evident nuclear localization signal (NLS) of <i>G. lamblia</i> MYB2 (GlMYB2) has not been defined. Based on putative GlMYB2 NLSs predicted by 2 programs, a series of plasmids expressing hemagglutinin (HA)-tagged GlMYB2 from the promoter of <i>G. lamblia</i> glutamate dehydrogenase were constructed and transfected into Giardia trophozoites. Immunofluorescence assays using anti-HA antibodies indicated that GlMYB2 amino acid sequence #507–#530 was required for the nuclear localization of GlMYB2, and this sequence was named as NLS<sub>GlMYB2</sub>. We further verified this finding by demonstrating the nuclear location of a protein obtained by the fusion of NLS<sub>GlMYB2</sub> and <i>G. lamblia</i> glyceraldehyde 3-phosphate dehydrogenase, a non-nuclear protein. Our data on GlMYB2 will expand our understanding on NLSs functioning in <i>G. lamblia.</i>