A Novel Spo11 Homologue Functions as a Positive Regulator in Cyst Differentiation in <i>Giardia lamblia</i>

<i>Giardia lamblia</i> persists in a dormant state with a protective cyst wall for transmission. It is incompletely known how three cyst wall proteins (CWPs) are coordinately synthesized during encystation. Meiotic recombination is required for sexual reproduction in animals, fungi, and...

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Autores principales:	Yu-Chien Chen, Szu-Yu Tung, Chia-Wei Huang, Soo-Wah Gan, Bo-Chi Lin, Chia-Wei Chen, Zi-Qi Lin, Chin-Hung Sun
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	cyst Spo11 transcription <i>Giardia</i> differentiation DNA binding protein Biology (General) QH301-705.5 Chemistry QD1-999
Acceso en línea:	https://doaj.org/article/ec9fff1f402d4999bbf90c6ad3f8b2a0
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id	oai:doaj.org-article:ec9fff1f402d4999bbf90c6ad3f8b2a0
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spelling	oai:doaj.org-article:ec9fff1f402d4999bbf90c6ad3f8b2a02021-11-11T17:19:02ZA Novel Spo11 Homologue Functions as a Positive Regulator in Cyst Differentiation in <i>Giardia lamblia</i>10.3390/ijms2221119021422-00671661-6596https://doaj.org/article/ec9fff1f402d4999bbf90c6ad3f8b2a02021-11-01T00:00:00Zhttps://www.mdpi.com/1422-0067/22/21/11902https://doaj.org/toc/1661-6596https://doaj.org/toc/1422-0067<i>Giardia lamblia</i> persists in a dormant state with a protective cyst wall for transmission. It is incompletely known how three cyst wall proteins (CWPs) are coordinately synthesized during encystation. Meiotic recombination is required for sexual reproduction in animals, fungi, and plants. It is initiated by formation of double-stranded breaks by a topoisomerase-like Spo11. It has been shown that exchange of genetic material in the fused nuclei occurs during <i>Giardia</i> encystation, suggesting parasexual recombination processes of this protozoan. <i>Giardia</i> possesses an evolutionarily conserved Spo11 with typical domains for cleavage reaction and an upregulated expression pattern during encystation. In this study, we asked whether Spo11 can activate encystation process, like other topoisomerases we previously characterized. We found that Spo11 was capable of binding to both single-stranded and double-stranded DNA in vitro and that it could also bind to the <i>cwp</i> promoters in vivo as accessed in chromatin immunoprecipitation assays. Spo11 interacted with WRKY and MYB2 (named from myeloblastosis), transcription factors that can activate <i>cwp</i> gene expression during encystation. Interestingly, overexpression of Spo11 resulted in increased expression of <i>cwp1</i>-<i>3</i> and <i>myb2</i> genes and cyst formation. Mutation of the Tyr residue for the active site or two conserved residues corresponding to key DNA-binding residues for <i>Arabidopsis</i> Spo11 reduced the levels of <i>cwp1-3</i> and <i>myb2</i> gene expression and cyst formation. Targeted disruption of <i>spo11</i> gene with CRISPR/Cas9 system led to a significant decrease in <i>cwp1-3</i> and <i>myb2</i> gene expression and cyst number. Our results suggest that Spo11 acts as a positive regulator for <i>Giardia</i> differentiation into cyst.Yu-Chien ChenSzu-Yu TungChia-Wei HuangSoo-Wah GanBo-Chi LinChia-Wei ChenZi-Qi LinChin-Hung SunMDPI AGarticlecystSpo11transcription<i>Giardia</i>differentiationDNA binding proteinBiology (General)QH301-705.5ChemistryQD1-999ENInternational Journal of Molecular Sciences, Vol 22, Iss 11902, p 11902 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	cyst Spo11 transcription <i>Giardia</i> differentiation DNA binding protein Biology (General) QH301-705.5 Chemistry QD1-999
spellingShingle	cyst Spo11 transcription <i>Giardia</i> differentiation DNA binding protein Biology (General) QH301-705.5 Chemistry QD1-999 Yu-Chien Chen Szu-Yu Tung Chia-Wei Huang Soo-Wah Gan Bo-Chi Lin Chia-Wei Chen Zi-Qi Lin Chin-Hung Sun A Novel Spo11 Homologue Functions as a Positive Regulator in Cyst Differentiation in <i>Giardia lamblia</i>
description	<i>Giardia lamblia</i> persists in a dormant state with a protective cyst wall for transmission. It is incompletely known how three cyst wall proteins (CWPs) are coordinately synthesized during encystation. Meiotic recombination is required for sexual reproduction in animals, fungi, and plants. It is initiated by formation of double-stranded breaks by a topoisomerase-like Spo11. It has been shown that exchange of genetic material in the fused nuclei occurs during <i>Giardia</i> encystation, suggesting parasexual recombination processes of this protozoan. <i>Giardia</i> possesses an evolutionarily conserved Spo11 with typical domains for cleavage reaction and an upregulated expression pattern during encystation. In this study, we asked whether Spo11 can activate encystation process, like other topoisomerases we previously characterized. We found that Spo11 was capable of binding to both single-stranded and double-stranded DNA in vitro and that it could also bind to the <i>cwp</i> promoters in vivo as accessed in chromatin immunoprecipitation assays. Spo11 interacted with WRKY and MYB2 (named from myeloblastosis), transcription factors that can activate <i>cwp</i> gene expression during encystation. Interestingly, overexpression of Spo11 resulted in increased expression of <i>cwp1</i>-<i>3</i> and <i>myb2</i> genes and cyst formation. Mutation of the Tyr residue for the active site or two conserved residues corresponding to key DNA-binding residues for <i>Arabidopsis</i> Spo11 reduced the levels of <i>cwp1-3</i> and <i>myb2</i> gene expression and cyst formation. Targeted disruption of <i>spo11</i> gene with CRISPR/Cas9 system led to a significant decrease in <i>cwp1-3</i> and <i>myb2</i> gene expression and cyst number. Our results suggest that Spo11 acts as a positive regulator for <i>Giardia</i> differentiation into cyst.
format	article
author	Yu-Chien Chen Szu-Yu Tung Chia-Wei Huang Soo-Wah Gan Bo-Chi Lin Chia-Wei Chen Zi-Qi Lin Chin-Hung Sun
author_facet	Yu-Chien Chen Szu-Yu Tung Chia-Wei Huang Soo-Wah Gan Bo-Chi Lin Chia-Wei Chen Zi-Qi Lin Chin-Hung Sun
author_sort	Yu-Chien Chen
title	A Novel Spo11 Homologue Functions as a Positive Regulator in Cyst Differentiation in <i>Giardia lamblia</i>
title_short	A Novel Spo11 Homologue Functions as a Positive Regulator in Cyst Differentiation in <i>Giardia lamblia</i>
title_full	A Novel Spo11 Homologue Functions as a Positive Regulator in Cyst Differentiation in <i>Giardia lamblia</i>
title_fullStr	A Novel Spo11 Homologue Functions as a Positive Regulator in Cyst Differentiation in <i>Giardia lamblia</i>
title_full_unstemmed	A Novel Spo11 Homologue Functions as a Positive Regulator in Cyst Differentiation in <i>Giardia lamblia</i>
title_sort	novel spo11 homologue functions as a positive regulator in cyst differentiation in <i>giardia lamblia</i>
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/ec9fff1f402d4999bbf90c6ad3f8b2a0
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A Novel Spo11 Homologue Functions as a Positive Regulator in Cyst Differentiation in <i>Giardia lamblia</i>

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