A Novel Multiprotein Bridging Factor 1-Like Protein Induces Cyst Wall Protein Gene Expression and Cyst Differentiation in <i>Giardia lamblia</i>
The capacity to synthesize a protective cyst wall is critical for infectivity of <i>Giardia lamblia</i>. It is of interest to know the mechanism of coordinated synthesis of three cyst wall proteins (CWPs) during encystation, a differentiation process. Multiprotein bridging factor 1 (MBF1...
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MDPI AG
2021-01-01
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| Series: | International Journal of Molecular Sciences |
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| author | Shao-Wei Huang Zi-Qi Lin Szu-Yu Tung Li-Hsin Su Chun-Che Ho Gilbert Aaron Lee Chin-Hung Sun |
| author_facet | Shao-Wei Huang Zi-Qi Lin Szu-Yu Tung Li-Hsin Su Chun-Che Ho Gilbert Aaron Lee Chin-Hung Sun |
| author_sort | Shao-Wei Huang |
| collection | DOAJ |
| description | The capacity to synthesize a protective cyst wall is critical for infectivity of <i>Giardia lamblia</i>. It is of interest to know the mechanism of coordinated synthesis of three cyst wall proteins (CWPs) during encystation, a differentiation process. Multiprotein bridging factor 1 (MBF1) gene family is a group of transcription coactivators that bridge various transcription factors. They are involved in cell growth and differentiation in yeast and animals, or in stress response in fungi and plants. We asked whether <i>Giardia</i> has MBF1-like genes and whether their products influence gene expression. BLAST searches of the <i>Giardia</i> genome database identified one gene encoding a putative MBF1 protein with a helix-turn-helix domain. We found that it can specifically bind to the AT-rich initiator promoters of the encystation-induced cwp1-3 and myb2 genes. MBF1 localized to cell nuclei and cytoplasm with higher expression during encystation. In addition, overexpression of MBF1 induced cwp1-3 and myb2 gene expression and cyst generation. Mutation of the helixes in the helix-turn-helix domain reduced cwp1-3 and myb2 gene expression and cyst generation. Chromatin immunoprecipitation assays confirmed the binding of MBF1 to the promoters with its binding sites in vivo. We also found that MBF1 can interact with E2F1, Pax2, WRKY, and Myb2 transcription factors that coordinately up-regulate the cwp genes during encystation. Using a CRISPR/Cas9 system for targeted disruption of mbf1 gene, we found a downregulation of cwp1-3 and myb2 genes and decrease of cyst generation. Our results suggest that MBF1 is functionally conserved and positively regulates <i>Giardia</i> cyst differentiation. |
| first_indexed | 2024-03-09T03:16:55Z |
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| language | English |
| last_indexed | 2024-03-09T03:16:55Z |
| publishDate | 2021-01-01 |
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| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-5acd08738b4743249241094c0bf0bbeb2023-12-03T15:16:50ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672021-01-01223137010.3390/ijms22031370A Novel Multiprotein Bridging Factor 1-Like Protein Induces Cyst Wall Protein Gene Expression and Cyst Differentiation in <i>Giardia lamblia</i>Shao-Wei Huang0Zi-Qi Lin1Szu-Yu Tung2Li-Hsin Su3Chun-Che Ho4Gilbert Aaron Lee5Chin-Hung Sun6Department of Tropical Medicine and Parasitology, College of Medicine, National Taiwan University, Taipei 100, TaiwanDepartment of Tropical Medicine and Parasitology, College of Medicine, National Taiwan University, Taipei 100, TaiwanDepartment of Tropical Medicine and Parasitology, College of Medicine, National Taiwan University, Taipei 100, TaiwanDepartment of Tropical Medicine and Parasitology, College of Medicine, National Taiwan University, Taipei 100, TaiwanDepartment of Tropical Medicine and Parasitology, College of Medicine, National Taiwan University, Taipei 100, TaiwanDepartment of Medical Research, Taipei Medical University Hospital, Taipei 110, TaiwanDepartment of Tropical Medicine and Parasitology, College of Medicine, National Taiwan University, Taipei 100, TaiwanThe capacity to synthesize a protective cyst wall is critical for infectivity of <i>Giardia lamblia</i>. It is of interest to know the mechanism of coordinated synthesis of three cyst wall proteins (CWPs) during encystation, a differentiation process. Multiprotein bridging factor 1 (MBF1) gene family is a group of transcription coactivators that bridge various transcription factors. They are involved in cell growth and differentiation in yeast and animals, or in stress response in fungi and plants. We asked whether <i>Giardia</i> has MBF1-like genes and whether their products influence gene expression. BLAST searches of the <i>Giardia</i> genome database identified one gene encoding a putative MBF1 protein with a helix-turn-helix domain. We found that it can specifically bind to the AT-rich initiator promoters of the encystation-induced cwp1-3 and myb2 genes. MBF1 localized to cell nuclei and cytoplasm with higher expression during encystation. In addition, overexpression of MBF1 induced cwp1-3 and myb2 gene expression and cyst generation. Mutation of the helixes in the helix-turn-helix domain reduced cwp1-3 and myb2 gene expression and cyst generation. Chromatin immunoprecipitation assays confirmed the binding of MBF1 to the promoters with its binding sites in vivo. We also found that MBF1 can interact with E2F1, Pax2, WRKY, and Myb2 transcription factors that coordinately up-regulate the cwp genes during encystation. Using a CRISPR/Cas9 system for targeted disruption of mbf1 gene, we found a downregulation of cwp1-3 and myb2 genes and decrease of cyst generation. Our results suggest that MBF1 is functionally conserved and positively regulates <i>Giardia</i> cyst differentiation.https://www.mdpi.com/1422-0067/22/3/1370cystMBF1transcription<i>Giardia</i>differentiationDNA binding protein |
| spellingShingle | Shao-Wei Huang Zi-Qi Lin Szu-Yu Tung Li-Hsin Su Chun-Che Ho Gilbert Aaron Lee Chin-Hung Sun A Novel Multiprotein Bridging Factor 1-Like Protein Induces Cyst Wall Protein Gene Expression and Cyst Differentiation in <i>Giardia lamblia</i> International Journal of Molecular Sciences cyst MBF1 transcription <i>Giardia</i> differentiation DNA binding protein |
| title | A Novel Multiprotein Bridging Factor 1-Like Protein Induces Cyst Wall Protein Gene Expression and Cyst Differentiation in <i>Giardia lamblia</i> |
| title_full | A Novel Multiprotein Bridging Factor 1-Like Protein Induces Cyst Wall Protein Gene Expression and Cyst Differentiation in <i>Giardia lamblia</i> |
| title_fullStr | A Novel Multiprotein Bridging Factor 1-Like Protein Induces Cyst Wall Protein Gene Expression and Cyst Differentiation in <i>Giardia lamblia</i> |
| title_full_unstemmed | A Novel Multiprotein Bridging Factor 1-Like Protein Induces Cyst Wall Protein Gene Expression and Cyst Differentiation in <i>Giardia lamblia</i> |
| title_short | A Novel Multiprotein Bridging Factor 1-Like Protein Induces Cyst Wall Protein Gene Expression and Cyst Differentiation in <i>Giardia lamblia</i> |
| title_sort | novel multiprotein bridging factor 1 like protein induces cyst wall protein gene expression and cyst differentiation in i giardia lamblia i |
| topic | cyst MBF1 transcription <i>Giardia</i> differentiation DNA binding protein |
| url | https://www.mdpi.com/1422-0067/22/3/1370 |
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