The Integrity of the Cell Wall and Its Remodeling during Heterocyst Differentiation Are Regulated by Phylogenetically Conserved Small RNA Yfr1 in <italic toggle="yes">Nostoc</italic> sp. Strain PCC 7120

ABSTRACT Yfr1 is a strictly conserved small RNA in cyanobacteria. A bioinformatic prediction to identify possible interactions of Yfr1 with mRNAs was carried out by using the sequences of Yfr1 from several heterocyst-forming strains, including Nostoc sp. strain PCC 7120. The results of the predictio...

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Autores principales:	Manuel Brenes-Álvarez, Agustín Vioque, Alicia M. Muro-Pastor
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2020
Materias:	Anabaena cyanobacteria heterocyst differentiation regulatory RNAs small RNAs Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/86d7d1489ca4456cb56bdb02ef24332a
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spelling	oai:doaj.org-article:86d7d1489ca4456cb56bdb02ef24332a2021-11-15T15:56:57ZThe Integrity of the Cell Wall and Its Remodeling during Heterocyst Differentiation Are Regulated by Phylogenetically Conserved Small RNA Yfr1 in <italic toggle="yes">Nostoc</italic> sp. Strain PCC 712010.1128/mBio.02599-192150-7511https://doaj.org/article/86d7d1489ca4456cb56bdb02ef24332a2020-02-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.02599-19https://doaj.org/toc/2150-7511ABSTRACT Yfr1 is a strictly conserved small RNA in cyanobacteria. A bioinformatic prediction to identify possible interactions of Yfr1 with mRNAs was carried out by using the sequences of Yfr1 from several heterocyst-forming strains, including Nostoc sp. strain PCC 7120. The results of the prediction were enriched in genes encoding outer membrane proteins and enzymes related to peptidoglycan biosynthesis and turnover. Heterologous expression assays with Escherichia coli demonstrated direct interactions of Yfr1 with mRNAs of 11 of the candidate genes. The expression of 10 of them (alr2458, alr4550, murC, all4829, all2158, mraY, alr2269, alr0834, conR, patN) was repressed by interaction with Yfr1, whereas the expression of amiC2, encoding an amidase, was increased. The interactions between Yfr1 and the 11 mRNAs were confirmed by site-directed mutagenesis of Yfr1. Furthermore, a Nostoc strain with reduced levels of Yfr1 had larger amounts of mraY and murC mRNAs, supporting a role for Yfr1 in the regulation of those genes. Nostoc strains with either reduced or increased expression of Yfr1 showed anomalies in cell wall completion and were more sensitive to vancomycin than the wild-type strain. Furthermore, growth in the absence of combined nitrogen, which involves the differentiation of heterocysts, was compromised in the strain overexpressing Yfr1, and filaments were broken at the connections between vegetative cells and heterocysts. These results indicate that Yfr1 is an important regulator of cell wall homeostasis and correct cell wall remodeling during heterocyst differentiation. IMPORTANCE Bacterial small RNAs (sRNAs) are important players affecting the regulation of essentially every aspect of bacterial physiology. The cell wall is a highly dynamic structure that protects bacteria from their fluctuating environment. Cell envelope remodeling is particularly critical for bacteria that undergo differentiation processes, such as spore formation or differentiation of heterocysts. Heterocyst development involves the deposition of additional layers of glycolipids and polysaccharides outside the outer membrane. Here, we show that a cyanobacterial phylogenetically conserved small regulatory RNA, Yfr1, coordinates the expression of proteins involved in cell wall-related processes, including peptidoglycan metabolism and transport of different molecules, as well as expression of several proteins involved in heterocyst differentiation.Manuel Brenes-ÁlvarezAgustín VioqueAlicia M. Muro-PastorAmerican Society for MicrobiologyarticleAnabaenacyanobacteriaheterocyst differentiationregulatory RNAssmall RNAsMicrobiologyQR1-502ENmBio, Vol 11, Iss 1 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	Anabaena cyanobacteria heterocyst differentiation regulatory RNAs small RNAs Microbiology QR1-502
spellingShingle	Anabaena cyanobacteria heterocyst differentiation regulatory RNAs small RNAs Microbiology QR1-502 Manuel Brenes-Álvarez Agustín Vioque Alicia M. Muro-Pastor The Integrity of the Cell Wall and Its Remodeling during Heterocyst Differentiation Are Regulated by Phylogenetically Conserved Small RNA Yfr1 in <italic toggle="yes">Nostoc</italic> sp. Strain PCC 7120
description	ABSTRACT Yfr1 is a strictly conserved small RNA in cyanobacteria. A bioinformatic prediction to identify possible interactions of Yfr1 with mRNAs was carried out by using the sequences of Yfr1 from several heterocyst-forming strains, including Nostoc sp. strain PCC 7120. The results of the prediction were enriched in genes encoding outer membrane proteins and enzymes related to peptidoglycan biosynthesis and turnover. Heterologous expression assays with Escherichia coli demonstrated direct interactions of Yfr1 with mRNAs of 11 of the candidate genes. The expression of 10 of them (alr2458, alr4550, murC, all4829, all2158, mraY, alr2269, alr0834, conR, patN) was repressed by interaction with Yfr1, whereas the expression of amiC2, encoding an amidase, was increased. The interactions between Yfr1 and the 11 mRNAs were confirmed by site-directed mutagenesis of Yfr1. Furthermore, a Nostoc strain with reduced levels of Yfr1 had larger amounts of mraY and murC mRNAs, supporting a role for Yfr1 in the regulation of those genes. Nostoc strains with either reduced or increased expression of Yfr1 showed anomalies in cell wall completion and were more sensitive to vancomycin than the wild-type strain. Furthermore, growth in the absence of combined nitrogen, which involves the differentiation of heterocysts, was compromised in the strain overexpressing Yfr1, and filaments were broken at the connections between vegetative cells and heterocysts. These results indicate that Yfr1 is an important regulator of cell wall homeostasis and correct cell wall remodeling during heterocyst differentiation. IMPORTANCE Bacterial small RNAs (sRNAs) are important players affecting the regulation of essentially every aspect of bacterial physiology. The cell wall is a highly dynamic structure that protects bacteria from their fluctuating environment. Cell envelope remodeling is particularly critical for bacteria that undergo differentiation processes, such as spore formation or differentiation of heterocysts. Heterocyst development involves the deposition of additional layers of glycolipids and polysaccharides outside the outer membrane. Here, we show that a cyanobacterial phylogenetically conserved small regulatory RNA, Yfr1, coordinates the expression of proteins involved in cell wall-related processes, including peptidoglycan metabolism and transport of different molecules, as well as expression of several proteins involved in heterocyst differentiation.
format	article
author	Manuel Brenes-Álvarez Agustín Vioque Alicia M. Muro-Pastor
author_facet	Manuel Brenes-Álvarez Agustín Vioque Alicia M. Muro-Pastor
author_sort	Manuel Brenes-Álvarez
title	The Integrity of the Cell Wall and Its Remodeling during Heterocyst Differentiation Are Regulated by Phylogenetically Conserved Small RNA Yfr1 in <italic toggle="yes">Nostoc</italic> sp. Strain PCC 7120
title_short	The Integrity of the Cell Wall and Its Remodeling during Heterocyst Differentiation Are Regulated by Phylogenetically Conserved Small RNA Yfr1 in <italic toggle="yes">Nostoc</italic> sp. Strain PCC 7120
title_full	The Integrity of the Cell Wall and Its Remodeling during Heterocyst Differentiation Are Regulated by Phylogenetically Conserved Small RNA Yfr1 in <italic toggle="yes">Nostoc</italic> sp. Strain PCC 7120
title_fullStr	The Integrity of the Cell Wall and Its Remodeling during Heterocyst Differentiation Are Regulated by Phylogenetically Conserved Small RNA Yfr1 in <italic toggle="yes">Nostoc</italic> sp. Strain PCC 7120
title_full_unstemmed	The Integrity of the Cell Wall and Its Remodeling during Heterocyst Differentiation Are Regulated by Phylogenetically Conserved Small RNA Yfr1 in <italic toggle="yes">Nostoc</italic> sp. Strain PCC 7120
title_sort	integrity of the cell wall and its remodeling during heterocyst differentiation are regulated by phylogenetically conserved small rna yfr1 in <italic toggle="yes">nostoc</italic> sp. strain pcc 7120
publisher	American Society for Microbiology
publishDate	2020
url	https://doaj.org/article/86d7d1489ca4456cb56bdb02ef24332a
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The Integrity of the Cell Wall and Its Remodeling during Heterocyst Differentiation Are Regulated by Phylogenetically Conserved Small RNA Yfr1 in <italic toggle="yes">Nostoc</italic> sp. Strain PCC 7120

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