Universität zu Köln

Melior, Hendrik, Maass, Sandra, Li, Siqi, Foerstner, Konrad U., Azarderakhsh, Saina, Varadarajan, Adithi R., Stoetzel, Maximilian, Elhossary, Muhammad, Barth-Weber, Susanne, Ahrens, Christian H. ORCID: 0000-0002-8148-7257, Becher, Doerte and Evguenieva-Hackenberg, Elena (2020). The Leader Peptide peTrpL Forms Antibiotic-Containing Ribonucleoprotein Complexes for Posttranscriptional Regulation of Multiresistance Genes. mBio, 11 (3). WASHINGTON: AMER SOC MICROBIOLOGY. ISSN 2150-7511

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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 tryptophanindependent 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 Aiphaproteobacteria. 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.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Melior, Hendrik UNSPECIFIED UNSPECIFIED UNSPECIFIED Maass, Sandra UNSPECIFIED UNSPECIFIED UNSPECIFIED Li, Siqi UNSPECIFIED UNSPECIFIED UNSPECIFIED Foerstner, Konrad U. UNSPECIFIED UNSPECIFIED UNSPECIFIED Azarderakhsh, Saina UNSPECIFIED UNSPECIFIED UNSPECIFIED Varadarajan, Adithi R. UNSPECIFIED UNSPECIFIED UNSPECIFIED Stoetzel, Maximilian UNSPECIFIED UNSPECIFIED UNSPECIFIED Elhossary, Muhammad UNSPECIFIED UNSPECIFIED UNSPECIFIED Barth-Weber, Susanne UNSPECIFIED UNSPECIFIED UNSPECIFIED Ahrens, Christian H. UNSPECIFIED orcid.org/0000-0002-8148-7257 UNSPECIFIED Becher, Doerte UNSPECIFIED UNSPECIFIED UNSPECIFIED Evguenieva-Hackenberg, Elena UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-334635
DOI:	10.1128/mBio.01027-20
Journal or Publication Title:	mBio
Volume:	11
Number:	3
Date:	2020
Publisher:	AMER SOC MICROBIOLOGY
Place of Publication:	WASHINGTON
ISSN:	2150-7511
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language MULTIDRUG EFFLUX SYSTEM; MESSENGER-RNA; TRANSCRIPTION ATTENUATION; PHOTOSYNTHESIS GENES; COMPARATIVE GENOMICS; CRYSTAL-STRUCTURES; RESISTANCE; PROTEIN; EXPRESSION; ACRB Multiple languages Microbiology Multiple languages
URI:	http://kups.ub.uni-koeln.de/id/eprint/33463