Functional Identification of a Nuclear Localization Signal of MYB2 Protein in Giardia lamblia
2
조회
0
다운로드
0
인용
0
참고문헌
키워드
초록
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 pro- grams, 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.
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 pro- grams, 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.
참고문헌 (10)
- Yang H, "Identification of an encystation-specific transcription factor, Myb protein in Giardia lamblia", Mol Biochem Parasitol, 128, pp.167~174 (2003)[DOI]
- Sun CH, "A novel Myb-related protein involved in transcriptional activation of encystation genes in Giardia lamblia", Mol Microbiol, 46, pp.971~984 (2002)[DOI]
- Lange A, "Classical nuclear localization signals : definition, function, and interaction with importin α", J Biol Chem, 282, pp.5101~5105 (2007)[DOI]
- Hagen KD, "Novel structural components of the ventral disc and lateral crest in Giardia intestinalis", PLoS Negl Trop Dis, 5, pp.e1442~ (2011)[DOI]
- Nguyen Ba AN, "NLStradamus : a simple hidden Markov model for nuclear localization signal prediction", BMC Bioinformatics, 10, pp.202~ (2009)[DOI]
- McInally SG, "Robust and stable transcriptional repression in Giardia using CRISPRi", Mol Biol Cell, 30, pp.119~130 (2019)[DOI]
- Gourguechon S, "Rapid tagging and integration of genes in Giardia intestinalis", Eukary Cell, 10, pp.142~145 (2011)[DOI]
- Keister DB, "Axenic culture of Giardia lamblia in TYI-S-33 medium supplemented with bile", Trans R Soc Trop Med Hyg, 77, pp.487~488 (1983)[DOI]
- Marfori M, "Molecular basis for specificity of nuclear import and prediction of nuclear localization", Biochim Biophy Acta, 1813, pp.1562~1577 (2011)[DOI]
- Canela-Pérez I, "Nuclear localization signals in trypanosomal proteins", Mol Biochem Parasitol, 229, pp.15~23 (2019)[DOI]
