Functional Identification of a Nuclear Localization Signal of MYB2 Protein in Giardia lamblia

Korean J Parasitol. 2020 Dec;58(6):675-679. doi: 10.3347/kjp.2020.58.6.675. Epub 2020 Dec 29.

Abstract

MYB2 protein was identified as a transcription factor that showed encystation-induced expression in Giardia lamblia. Although nuclear import is essential for the functioning of a transcription factor, an evident nuclear localization signal (NLS) of G. lamblia 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 G. lamblia 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 NLSGlMYB2. We further verified this finding by demonstrating the nuclear location of a protein obtained by the fusion of NLSGlMYB2 and G. lamblia glyceraldehyde 3-phosphate dehydrogenase, a non-nuclear protein. Our data on GlMYB2 will expand our understanding on NLSs functioning in G. lamblia.

Keywords: Giardia lamblia; GlMYB2; nuclear localization signal.

MeSH terms

  • Amino Acid Sequence
  • Gene Expression Regulation, Developmental / genetics*
  • Gene Expression*
  • Giardia lamblia / enzymology
  • Giardia lamblia / growth & development*
  • Giardia lamblia / physiology*
  • Glutamate Dehydrogenase
  • Glyceraldehyde 3-Phosphate
  • Hemagglutinins
  • Nuclear Localization Signals / genetics*
  • Nuclear Localization Signals / metabolism*
  • Parasite Encystment / genetics*
  • Trans-Activators / chemistry
  • Trans-Activators / genetics*
  • Trans-Activators / metabolism*

Substances

  • Hemagglutinins
  • Myb2 protein, Giardia lamblia
  • Nuclear Localization Signals
  • Trans-Activators
  • Glyceraldehyde 3-Phosphate
  • Glutamate Dehydrogenase