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A Novel Splice Variant of the Masculinizing Gene Masc with piRNA-Cleavage-Site Defect Functions in Female External Genital Development in the Silkworm, Bombyx mori

In the silkworm, the sex-determination primary signal Fem controls sex differentiation by specific binding of Fem-derived piRNA to the cleavage site in Masc mRNA, thus inhibiting Masc protein production in the female. In this study, we identified a novel splicing isoform of Masc, named Masc-S, which...

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Autores principales: Zhao, Qin, Li, Juan, Wen, Mao-Yu, Wang, He, Wang, Yao, Wang, Kai-Xuan, Wan, Qiu-Xing, Zha, Xing-Fu
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6723575/
https://www.ncbi.nlm.nih.gov/pubmed/31366115
http://dx.doi.org/10.3390/biom9080318
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author Zhao, Qin
Li, Juan
Wen, Mao-Yu
Wang, He
Wang, Yao
Wang, Kai-Xuan
Wan, Qiu-Xing
Zha, Xing-Fu
author_facet Zhao, Qin
Li, Juan
Wen, Mao-Yu
Wang, He
Wang, Yao
Wang, Kai-Xuan
Wan, Qiu-Xing
Zha, Xing-Fu
author_sort Zhao, Qin
collection PubMed
description In the silkworm, the sex-determination primary signal Fem controls sex differentiation by specific binding of Fem-derived piRNA to the cleavage site in Masc mRNA, thus inhibiting Masc protein production in the female. In this study, we identified a novel splicing isoform of Masc, named Masc-S, which lacks the intact sequence of the cleavage site, encoding a C-terminal truncated protein. Results of RT-PCR showed that Masc-S was expressed in both sexes. Over-expression of Masc-S and Masc in female-specific cell lines showed that Masc-S could be translated against the Fem-piRNA cut. By RNA-protein pull-down, LC/MS/MS, and EMSA, we identified a protein BmEXU that specifically binds to an exclusive RNA sequence in Masc compared to Masc-S. Knockdown of Masc-S resulted in abnormal morphology in female external genital and increased expression of the Hox gene Abd-B, which similarly occurred by Bmexu RNAi. These results suggest that the splice variant Masc-S against Fem-piRNA plays an important role in female external genital development, of which function is opposite to that of full-length Masc. Our study provides new insights into the regulatory mechanism of sex determination in the silkworm.
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spelling pubmed-67235752019-09-10 A Novel Splice Variant of the Masculinizing Gene Masc with piRNA-Cleavage-Site Defect Functions in Female External Genital Development in the Silkworm, Bombyx mori Zhao, Qin Li, Juan Wen, Mao-Yu Wang, He Wang, Yao Wang, Kai-Xuan Wan, Qiu-Xing Zha, Xing-Fu Biomolecules Article In the silkworm, the sex-determination primary signal Fem controls sex differentiation by specific binding of Fem-derived piRNA to the cleavage site in Masc mRNA, thus inhibiting Masc protein production in the female. In this study, we identified a novel splicing isoform of Masc, named Masc-S, which lacks the intact sequence of the cleavage site, encoding a C-terminal truncated protein. Results of RT-PCR showed that Masc-S was expressed in both sexes. Over-expression of Masc-S and Masc in female-specific cell lines showed that Masc-S could be translated against the Fem-piRNA cut. By RNA-protein pull-down, LC/MS/MS, and EMSA, we identified a protein BmEXU that specifically binds to an exclusive RNA sequence in Masc compared to Masc-S. Knockdown of Masc-S resulted in abnormal morphology in female external genital and increased expression of the Hox gene Abd-B, which similarly occurred by Bmexu RNAi. These results suggest that the splice variant Masc-S against Fem-piRNA plays an important role in female external genital development, of which function is opposite to that of full-length Masc. Our study provides new insights into the regulatory mechanism of sex determination in the silkworm. MDPI 2019-07-30 /pmc/articles/PMC6723575/ /pubmed/31366115 http://dx.doi.org/10.3390/biom9080318 Text en © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Zhao, Qin
Li, Juan
Wen, Mao-Yu
Wang, He
Wang, Yao
Wang, Kai-Xuan
Wan, Qiu-Xing
Zha, Xing-Fu
A Novel Splice Variant of the Masculinizing Gene Masc with piRNA-Cleavage-Site Defect Functions in Female External Genital Development in the Silkworm, Bombyx mori
title A Novel Splice Variant of the Masculinizing Gene Masc with piRNA-Cleavage-Site Defect Functions in Female External Genital Development in the Silkworm, Bombyx mori
title_full A Novel Splice Variant of the Masculinizing Gene Masc with piRNA-Cleavage-Site Defect Functions in Female External Genital Development in the Silkworm, Bombyx mori
title_fullStr A Novel Splice Variant of the Masculinizing Gene Masc with piRNA-Cleavage-Site Defect Functions in Female External Genital Development in the Silkworm, Bombyx mori
title_full_unstemmed A Novel Splice Variant of the Masculinizing Gene Masc with piRNA-Cleavage-Site Defect Functions in Female External Genital Development in the Silkworm, Bombyx mori
title_short A Novel Splice Variant of the Masculinizing Gene Masc with piRNA-Cleavage-Site Defect Functions in Female External Genital Development in the Silkworm, Bombyx mori
title_sort novel splice variant of the masculinizing gene masc with pirna-cleavage-site defect functions in female external genital development in the silkworm, bombyx mori
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6723575/
https://www.ncbi.nlm.nih.gov/pubmed/31366115
http://dx.doi.org/10.3390/biom9080318
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