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Specific interaction of an RNA-binding protein with the 3′-UTR of its target mRNA is critical to oomycete sexual reproduction

Sexual reproduction is an essential stage of the oomycete life cycle. However, the functions of critical regulators in this biological process remain unclear due to a lack of genome editing technologies and functional genomic studies in oomycetes. The notorious oomycete pathogen Pythium ultimum is r...

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Autores principales: Feng, Hui, Wan, Chuanxu, Zhang, Zhichao, Chen, Han, Li, Zhipeng, Jiang, Haibin, Yin, Maozhu, Dong, Suomeng, Dou, Daolong, Wang, Yuanchao, Zheng, Xiaobo, Ye, Wenwu
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8547697/
https://www.ncbi.nlm.nih.gov/pubmed/34648596
http://dx.doi.org/10.1371/journal.ppat.1010001
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author Feng, Hui
Wan, Chuanxu
Zhang, Zhichao
Chen, Han
Li, Zhipeng
Jiang, Haibin
Yin, Maozhu
Dong, Suomeng
Dou, Daolong
Wang, Yuanchao
Zheng, Xiaobo
Ye, Wenwu
author_facet Feng, Hui
Wan, Chuanxu
Zhang, Zhichao
Chen, Han
Li, Zhipeng
Jiang, Haibin
Yin, Maozhu
Dong, Suomeng
Dou, Daolong
Wang, Yuanchao
Zheng, Xiaobo
Ye, Wenwu
author_sort Feng, Hui
collection PubMed
description Sexual reproduction is an essential stage of the oomycete life cycle. However, the functions of critical regulators in this biological process remain unclear due to a lack of genome editing technologies and functional genomic studies in oomycetes. The notorious oomycete pathogen Pythium ultimum is responsible for a variety of diseases in a broad range of plant species. In this study, we revealed the mechanism through which PuM90, a stage-specific Puf family RNA-binding protein, regulates oospore formation in P. ultimum. We developed the first CRISPR/Cas9 system-mediated gene knockout and in situ complementation methods for Pythium. PuM90-knockout mutants were significantly defective in oospore formation, with empty oogonia or oospores larger in size with thinner oospore walls compared with the wild type. A tripartite recognition motif (TRM) in the Puf domain of PuM90 could specifically bind to a UGUACAUA motif in the mRNA 3′ untranslated region (UTR) of PuFLP, which encodes a flavodoxin-like protein, and thereby repress PuFLP mRNA level to facilitate oospore formation. Phenotypes similar to PuM90-knockout mutants were observed with overexpression of PuFLP, mutation of key amino acids in the TRM of PuM90, or mutation of the 3′-UTR binding site in PuFLP. The results demonstrated that a specific interaction of the RNA-binding protein PuM90 with the 3′-UTR of PuFLP mRNA at the post-transcriptional regulation level is critical for the sexual reproduction of P. ultimum.
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spelling pubmed-85476972021-10-27 Specific interaction of an RNA-binding protein with the 3′-UTR of its target mRNA is critical to oomycete sexual reproduction Feng, Hui Wan, Chuanxu Zhang, Zhichao Chen, Han Li, Zhipeng Jiang, Haibin Yin, Maozhu Dong, Suomeng Dou, Daolong Wang, Yuanchao Zheng, Xiaobo Ye, Wenwu PLoS Pathog Research Article Sexual reproduction is an essential stage of the oomycete life cycle. However, the functions of critical regulators in this biological process remain unclear due to a lack of genome editing technologies and functional genomic studies in oomycetes. The notorious oomycete pathogen Pythium ultimum is responsible for a variety of diseases in a broad range of plant species. In this study, we revealed the mechanism through which PuM90, a stage-specific Puf family RNA-binding protein, regulates oospore formation in P. ultimum. We developed the first CRISPR/Cas9 system-mediated gene knockout and in situ complementation methods for Pythium. PuM90-knockout mutants were significantly defective in oospore formation, with empty oogonia or oospores larger in size with thinner oospore walls compared with the wild type. A tripartite recognition motif (TRM) in the Puf domain of PuM90 could specifically bind to a UGUACAUA motif in the mRNA 3′ untranslated region (UTR) of PuFLP, which encodes a flavodoxin-like protein, and thereby repress PuFLP mRNA level to facilitate oospore formation. Phenotypes similar to PuM90-knockout mutants were observed with overexpression of PuFLP, mutation of key amino acids in the TRM of PuM90, or mutation of the 3′-UTR binding site in PuFLP. The results demonstrated that a specific interaction of the RNA-binding protein PuM90 with the 3′-UTR of PuFLP mRNA at the post-transcriptional regulation level is critical for the sexual reproduction of P. ultimum. Public Library of Science 2021-10-14 /pmc/articles/PMC8547697/ /pubmed/34648596 http://dx.doi.org/10.1371/journal.ppat.1010001 Text en © 2021 Feng et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Feng, Hui
Wan, Chuanxu
Zhang, Zhichao
Chen, Han
Li, Zhipeng
Jiang, Haibin
Yin, Maozhu
Dong, Suomeng
Dou, Daolong
Wang, Yuanchao
Zheng, Xiaobo
Ye, Wenwu
Specific interaction of an RNA-binding protein with the 3′-UTR of its target mRNA is critical to oomycete sexual reproduction
title Specific interaction of an RNA-binding protein with the 3′-UTR of its target mRNA is critical to oomycete sexual reproduction
title_full Specific interaction of an RNA-binding protein with the 3′-UTR of its target mRNA is critical to oomycete sexual reproduction
title_fullStr Specific interaction of an RNA-binding protein with the 3′-UTR of its target mRNA is critical to oomycete sexual reproduction
title_full_unstemmed Specific interaction of an RNA-binding protein with the 3′-UTR of its target mRNA is critical to oomycete sexual reproduction
title_short Specific interaction of an RNA-binding protein with the 3′-UTR of its target mRNA is critical to oomycete sexual reproduction
title_sort specific interaction of an rna-binding protein with the 3′-utr of its target mrna is critical to oomycete sexual reproduction
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8547697/
https://www.ncbi.nlm.nih.gov/pubmed/34648596
http://dx.doi.org/10.1371/journal.ppat.1010001
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