The Leader Peptide peTrpL Forms Antibiotic-Containing Ribonucleoprotein Complexes for Posttranscriptional Regulation of Multiresistance Genes

ABSTRACT Bacterial ribosome-dependent attenuators are widespread posttranscriptional regulators. They harbor small upstream open reading frames (uORFs) encoding leader peptides, for which no functions in trans are known yet. In the plant symbiont Sinorhizobium meliloti, the tryptophan biosynthesis g...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Hendrik Melior, Sandra Maaß, Siqi Li, Konrad U. Förstner, Saina Azarderakhsh, Adithi R. Varadarajan, Maximilian Stötzel, Muhammad Elhossary, Susanne Barth-Weber, Christian H. Ahrens, Dörte Becher, Elena Evguenieva-Hackenberg
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2020
Materias:	antimicrobial compound leader peptide multidrug resistance nucleoprotein complex posttranscriptional regulation Agrobacterium tumefaciens Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/8fa148d0902848ea9801627da0c77e64
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:8fa148d0902848ea9801627da0c77e64
record_format	dspace
spelling	oai:doaj.org-article:8fa148d0902848ea9801627da0c77e642021-11-15T15:56:47ZThe Leader Peptide peTrpL Forms Antibiotic-Containing Ribonucleoprotein Complexes for Posttranscriptional Regulation of Multiresistance Genes10.1128/mBio.01027-202150-7511https://doaj.org/article/8fa148d0902848ea9801627da0c77e642020-06-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.01027-20https://doaj.org/toc/2150-7511ABSTRACT Bacterial ribosome-dependent attenuators are widespread posttranscriptional regulators. They harbor small upstream open reading frames (uORFs) encoding leader peptides, for which no functions in trans are known yet. In the plant symbiont Sinorhizobium meliloti, the tryptophan biosynthesis gene trpE(G) is preceded by the uORF trpL and is regulated by transcription attenuation according to tryptophan availability. However, trpLE(G) transcription is initiated independently of the tryptophan level in S. meliloti, thereby ensuring a largely tryptophan-independent production of the leader peptide peTrpL. Here, we provide evidence for a tryptophan-independent role of peTrpL in trans. We found that peTrpL increases the resistance toward tetracycline, erythromycin, chloramphenicol, and the flavonoid genistein, which are substrates of the major multidrug efflux pump SmeAB. Coimmunoprecipitation with a FLAG-peTrpL suggested smeR mRNA, which encodes the transcription repressor of smeABR, as a peptide target. Indeed, upon antibiotic exposure, smeR mRNA was destabilized and smeA stabilized in a peTrpL-dependent manner, showing that peTrpL acts in the differential regulation of smeABR. Furthermore, smeR mRNA was coimmunoprecipitated with peTrpL in antibiotic-dependent ribonucleoprotein (ARNP) complexes, which, in addition, contained an antibiotic-induced antisense RNA complementary to smeR. In vitro ARNP reconstitution revealed that the above-mentioned antibiotics and genistein directly support complex formation. A specific region of the antisense RNA was identified as a seed region for ARNP assembly in vitro. Altogether, our data show that peTrpL is involved in a mechanism for direct utilization of antimicrobial compounds in posttranscriptional regulation of multiresistance genes. Importantly, this role of peTrpL in resistance is conserved in other Alphaproteobacteria. IMPORTANCE Leader peptides encoded by transcription attenuators are widespread small proteins that are considered nonfunctional in trans. We found that the leader peptide peTrpL of the soil-dwelling plant symbiont Sinorhizobium meliloti is required for differential, posttranscriptional regulation of a multidrug resistance operon upon antibiotic exposure. Multiresistance achieved by efflux of different antimicrobial compounds ensures survival and competitiveness in nature and is important from both evolutionary and medical points of view. We show that the leader peptide forms antibiotic- and flavonoid-dependent ribonucleoprotein complexes (ARNPs) for destabilization of smeR mRNA encoding the transcription repressor of the major multidrug resistance operon. The seed region for ARNP assembly was localized in an antisense RNA, whose transcription is induced by antimicrobial compounds. The discovery of ARNP complexes as new players in multiresistance regulation opens new perspectives in understanding bacterial physiology and evolution and potentially provides new targets for antibacterial control.Hendrik MeliorSandra MaaßSiqi LiKonrad U. FörstnerSaina AzarderakhshAdithi R. VaradarajanMaximilian StötzelMuhammad ElhossarySusanne Barth-WeberChristian H. AhrensDörte BecherElena Evguenieva-HackenbergAmerican Society for Microbiologyarticleantimicrobial compoundleader peptidemultidrug resistancenucleoprotein complexposttranscriptional regulationAgrobacterium tumefaciensMicrobiologyQR1-502ENmBio, Vol 11, Iss 3 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	antimicrobial compound leader peptide multidrug resistance nucleoprotein complex posttranscriptional regulation Agrobacterium tumefaciens Microbiology QR1-502
spellingShingle	antimicrobial compound leader peptide multidrug resistance nucleoprotein complex posttranscriptional regulation Agrobacterium tumefaciens Microbiology QR1-502 Hendrik Melior Sandra Maaß Siqi Li Konrad U. Förstner Saina Azarderakhsh Adithi R. Varadarajan Maximilian Stötzel Muhammad Elhossary Susanne Barth-Weber Christian H. Ahrens Dörte Becher Elena Evguenieva-Hackenberg The Leader Peptide peTrpL Forms Antibiotic-Containing Ribonucleoprotein Complexes for Posttranscriptional Regulation of Multiresistance Genes
description	ABSTRACT Bacterial ribosome-dependent attenuators are widespread posttranscriptional regulators. They harbor small upstream open reading frames (uORFs) encoding leader peptides, for which no functions in trans are known yet. In the plant symbiont Sinorhizobium meliloti, the tryptophan biosynthesis gene trpE(G) is preceded by the uORF trpL and is regulated by transcription attenuation according to tryptophan availability. However, trpLE(G) transcription is initiated independently of the tryptophan level in S. meliloti, thereby ensuring a largely tryptophan-independent production of the leader peptide peTrpL. Here, we provide evidence for a tryptophan-independent role of peTrpL in trans. We found that peTrpL increases the resistance toward tetracycline, erythromycin, chloramphenicol, and the flavonoid genistein, which are substrates of the major multidrug efflux pump SmeAB. Coimmunoprecipitation with a FLAG-peTrpL suggested smeR mRNA, which encodes the transcription repressor of smeABR, as a peptide target. Indeed, upon antibiotic exposure, smeR mRNA was destabilized and smeA stabilized in a peTrpL-dependent manner, showing that peTrpL acts in the differential regulation of smeABR. Furthermore, smeR mRNA was coimmunoprecipitated with peTrpL in antibiotic-dependent ribonucleoprotein (ARNP) complexes, which, in addition, contained an antibiotic-induced antisense RNA complementary to smeR. In vitro ARNP reconstitution revealed that the above-mentioned antibiotics and genistein directly support complex formation. A specific region of the antisense RNA was identified as a seed region for ARNP assembly in vitro. Altogether, our data show that peTrpL is involved in a mechanism for direct utilization of antimicrobial compounds in posttranscriptional regulation of multiresistance genes. Importantly, this role of peTrpL in resistance is conserved in other Alphaproteobacteria. IMPORTANCE Leader peptides encoded by transcription attenuators are widespread small proteins that are considered nonfunctional in trans. We found that the leader peptide peTrpL of the soil-dwelling plant symbiont Sinorhizobium meliloti is required for differential, posttranscriptional regulation of a multidrug resistance operon upon antibiotic exposure. Multiresistance achieved by efflux of different antimicrobial compounds ensures survival and competitiveness in nature and is important from both evolutionary and medical points of view. We show that the leader peptide forms antibiotic- and flavonoid-dependent ribonucleoprotein complexes (ARNPs) for destabilization of smeR mRNA encoding the transcription repressor of the major multidrug resistance operon. The seed region for ARNP assembly was localized in an antisense RNA, whose transcription is induced by antimicrobial compounds. The discovery of ARNP complexes as new players in multiresistance regulation opens new perspectives in understanding bacterial physiology and evolution and potentially provides new targets for antibacterial control.
format	article
author	Hendrik Melior Sandra Maaß Siqi Li Konrad U. Förstner Saina Azarderakhsh Adithi R. Varadarajan Maximilian Stötzel Muhammad Elhossary Susanne Barth-Weber Christian H. Ahrens Dörte Becher Elena Evguenieva-Hackenberg
author_facet	Hendrik Melior Sandra Maaß Siqi Li Konrad U. Förstner Saina Azarderakhsh Adithi R. Varadarajan Maximilian Stötzel Muhammad Elhossary Susanne Barth-Weber Christian H. Ahrens Dörte Becher Elena Evguenieva-Hackenberg
author_sort	Hendrik Melior
title	The Leader Peptide peTrpL Forms Antibiotic-Containing Ribonucleoprotein Complexes for Posttranscriptional Regulation of Multiresistance Genes
title_short	The Leader Peptide peTrpL Forms Antibiotic-Containing Ribonucleoprotein Complexes for Posttranscriptional Regulation of Multiresistance Genes
title_full	The Leader Peptide peTrpL Forms Antibiotic-Containing Ribonucleoprotein Complexes for Posttranscriptional Regulation of Multiresistance Genes
title_fullStr	The Leader Peptide peTrpL Forms Antibiotic-Containing Ribonucleoprotein Complexes for Posttranscriptional Regulation of Multiresistance Genes
title_full_unstemmed	The Leader Peptide peTrpL Forms Antibiotic-Containing Ribonucleoprotein Complexes for Posttranscriptional Regulation of Multiresistance Genes
title_sort	leader peptide petrpl forms antibiotic-containing ribonucleoprotein complexes for posttranscriptional regulation of multiresistance genes
publisher	American Society for Microbiology
publishDate	2020
url	https://doaj.org/article/8fa148d0902848ea9801627da0c77e64
work_keys_str_mv	AT hendrikmelior theleaderpeptidepetrplformsantibioticcontainingribonucleoproteincomplexesforposttranscriptionalregulationofmultiresistancegenes AT sandramaaß theleaderpeptidepetrplformsantibioticcontainingribonucleoproteincomplexesforposttranscriptionalregulationofmultiresistancegenes AT siqili theleaderpeptidepetrplformsantibioticcontainingribonucleoproteincomplexesforposttranscriptionalregulationofmultiresistancegenes AT konraduforstner theleaderpeptidepetrplformsantibioticcontainingribonucleoproteincomplexesforposttranscriptionalregulationofmultiresistancegenes AT sainaazarderakhsh theleaderpeptidepetrplformsantibioticcontainingribonucleoproteincomplexesforposttranscriptionalregulationofmultiresistancegenes AT adithirvaradarajan theleaderpeptidepetrplformsantibioticcontainingribonucleoproteincomplexesforposttranscriptionalregulationofmultiresistancegenes AT maximilianstotzel theleaderpeptidepetrplformsantibioticcontainingribonucleoproteincomplexesforposttranscriptionalregulationofmultiresistancegenes AT muhammadelhossary theleaderpeptidepetrplformsantibioticcontainingribonucleoproteincomplexesforposttranscriptionalregulationofmultiresistancegenes AT susannebarthweber theleaderpeptidepetrplformsantibioticcontainingribonucleoproteincomplexesforposttranscriptionalregulationofmultiresistancegenes AT christianhahrens theleaderpeptidepetrplformsantibioticcontainingribonucleoproteincomplexesforposttranscriptionalregulationofmultiresistancegenes AT dortebecher theleaderpeptidepetrplformsantibioticcontainingribonucleoproteincomplexesforposttranscriptionalregulationofmultiresistancegenes AT elenaevguenievahackenberg theleaderpeptidepetrplformsantibioticcontainingribonucleoproteincomplexesforposttranscriptionalregulationofmultiresistancegenes AT hendrikmelior leaderpeptidepetrplformsantibioticcontainingribonucleoproteincomplexesforposttranscriptionalregulationofmultiresistancegenes AT sandramaaß leaderpeptidepetrplformsantibioticcontainingribonucleoproteincomplexesforposttranscriptionalregulationofmultiresistancegenes AT siqili leaderpeptidepetrplformsantibioticcontainingribonucleoproteincomplexesforposttranscriptionalregulationofmultiresistancegenes AT konraduforstner leaderpeptidepetrplformsantibioticcontainingribonucleoproteincomplexesforposttranscriptionalregulationofmultiresistancegenes AT sainaazarderakhsh leaderpeptidepetrplformsantibioticcontainingribonucleoproteincomplexesforposttranscriptionalregulationofmultiresistancegenes AT adithirvaradarajan leaderpeptidepetrplformsantibioticcontainingribonucleoproteincomplexesforposttranscriptionalregulationofmultiresistancegenes AT maximilianstotzel leaderpeptidepetrplformsantibioticcontainingribonucleoproteincomplexesforposttranscriptionalregulationofmultiresistancegenes AT muhammadelhossary leaderpeptidepetrplformsantibioticcontainingribonucleoproteincomplexesforposttranscriptionalregulationofmultiresistancegenes AT susannebarthweber leaderpeptidepetrplformsantibioticcontainingribonucleoproteincomplexesforposttranscriptionalregulationofmultiresistancegenes AT christianhahrens leaderpeptidepetrplformsantibioticcontainingribonucleoproteincomplexesforposttranscriptionalregulationofmultiresistancegenes AT dortebecher leaderpeptidepetrplformsantibioticcontainingribonucleoproteincomplexesforposttranscriptionalregulationofmultiresistancegenes AT elenaevguenievahackenberg leaderpeptidepetrplformsantibioticcontainingribonucleoproteincomplexesforposttranscriptionalregulationofmultiresistancegenes
_version_	1718427105069367296

The Leader Peptide peTrpL Forms Antibiotic-Containing Ribonucleoprotein Complexes for Posttranscriptional Regulation of Multiresistance Genes

Ejemplares similares