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DEADSouth protein localizes to germ plasm and is required for the development of primordial germ cells in Xenopus laevis

DEADSouth mRNA is a component of germ plasm in Xenopus laevis and encodes a DDX25 DEAD-box RNA helicase. To determine the intracellular localization of DEADSouth protein, we injected mRNA encoding DEADSouth tagged with mCherry fluorescent protein into fertilized eggs from transgenic Xenopus expressi...

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Autores principales: Yamaguchi, Takeshi, Taguchi, Ayaka, Watanabe, Kenji, Orii, Hidefumi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Company of Biologists 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3575653/
https://www.ncbi.nlm.nih.gov/pubmed/23429978
http://dx.doi.org/10.1242/bio.20123111
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author Yamaguchi, Takeshi
Taguchi, Ayaka
Watanabe, Kenji
Orii, Hidefumi
author_facet Yamaguchi, Takeshi
Taguchi, Ayaka
Watanabe, Kenji
Orii, Hidefumi
author_sort Yamaguchi, Takeshi
collection PubMed
description DEADSouth mRNA is a component of germ plasm in Xenopus laevis and encodes a DDX25 DEAD-box RNA helicase. To determine the intracellular localization of DEADSouth protein, we injected mRNA encoding DEADSouth tagged with mCherry fluorescent protein into fertilized eggs from transgenic Xenopus expressing EGFP fused with a mitochondrial targeting signal. The DEADSouth-mCherry fusion protein was localized to the germ plasm, a mitochondria-rich region in primordial germ cells (PGCs). DEADSouth overexpression resulted in a reduction of PGC numbers after stage 20. Conversely, DEADSouth knockdown using an antisense locked nucleic acid gapmer inhibited movement of the germ plasm from the cortex to the perinuclear region, resulting in inhibition of PGC division at stage 12 and a decrease in PGC numbers at later stages. The knockdown phenotype was rescued by intact DEADSouth mRNA, but not mutant mRNA encoding inactive DEADSouth helicase. Surprisingly, it was also rescued by mouse vasa homolog and Xenopus vasa-like gene 1 mRNAs that encode DDX4 RNA helicases. The rescue was dependent on the 3′ untranslated region (3′UTR) of DEADSouth mRNA, which was used for PGC-specific expression. The 3′UTR contributed to localization of the injected mRNA to the germ plasm, resulting in effective localization of DEADSouth protein. These results demonstrate that localization of DEADSouth helicase to the germ plasm is required for proper PGC development in Xenopus laevis.
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spelling pubmed-35756532013-02-21 DEADSouth protein localizes to germ plasm and is required for the development of primordial germ cells in Xenopus laevis Yamaguchi, Takeshi Taguchi, Ayaka Watanabe, Kenji Orii, Hidefumi Biol Open Research Article DEADSouth mRNA is a component of germ plasm in Xenopus laevis and encodes a DDX25 DEAD-box RNA helicase. To determine the intracellular localization of DEADSouth protein, we injected mRNA encoding DEADSouth tagged with mCherry fluorescent protein into fertilized eggs from transgenic Xenopus expressing EGFP fused with a mitochondrial targeting signal. The DEADSouth-mCherry fusion protein was localized to the germ plasm, a mitochondria-rich region in primordial germ cells (PGCs). DEADSouth overexpression resulted in a reduction of PGC numbers after stage 20. Conversely, DEADSouth knockdown using an antisense locked nucleic acid gapmer inhibited movement of the germ plasm from the cortex to the perinuclear region, resulting in inhibition of PGC division at stage 12 and a decrease in PGC numbers at later stages. The knockdown phenotype was rescued by intact DEADSouth mRNA, but not mutant mRNA encoding inactive DEADSouth helicase. Surprisingly, it was also rescued by mouse vasa homolog and Xenopus vasa-like gene 1 mRNAs that encode DDX4 RNA helicases. The rescue was dependent on the 3′ untranslated region (3′UTR) of DEADSouth mRNA, which was used for PGC-specific expression. The 3′UTR contributed to localization of the injected mRNA to the germ plasm, resulting in effective localization of DEADSouth protein. These results demonstrate that localization of DEADSouth helicase to the germ plasm is required for proper PGC development in Xenopus laevis. The Company of Biologists 2012-11-28 /pmc/articles/PMC3575653/ /pubmed/23429978 http://dx.doi.org/10.1242/bio.20123111 Text en © 2012. Published by The Company of Biologists Ltd http://creativecommons.org/licenses/by-nc-sa/3.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial Share Alike License (http://creativecommons.org/licenses/by-nc-sa/3.0/).
spellingShingle Research Article
Yamaguchi, Takeshi
Taguchi, Ayaka
Watanabe, Kenji
Orii, Hidefumi
DEADSouth protein localizes to germ plasm and is required for the development of primordial germ cells in Xenopus laevis
title DEADSouth protein localizes to germ plasm and is required for the development of primordial germ cells in Xenopus laevis
title_full DEADSouth protein localizes to germ plasm and is required for the development of primordial germ cells in Xenopus laevis
title_fullStr DEADSouth protein localizes to germ plasm and is required for the development of primordial germ cells in Xenopus laevis
title_full_unstemmed DEADSouth protein localizes to germ plasm and is required for the development of primordial germ cells in Xenopus laevis
title_short DEADSouth protein localizes to germ plasm and is required for the development of primordial germ cells in Xenopus laevis
title_sort deadsouth protein localizes to germ plasm and is required for the development of primordial germ cells in xenopus laevis
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3575653/
https://www.ncbi.nlm.nih.gov/pubmed/23429978
http://dx.doi.org/10.1242/bio.20123111
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