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An archaeal sRNA targeting cis- and trans-encoded mRNAs via two distinct domains

Please always quote using this URN: urn:nbn:de:bvb:20-opus-134972
  • We report on the characterization and target analysis of the small (s) RNA\(_{162}\) in the methanoarchaeon Methanosarcina mazei. Using a combination of genetic approaches, transcriptome analysis and computational predictions, the bicistronic MM2441-MM2440 mRNA encoding the transcription factor MM2441 and a protein of unknown function was identified as a potential target of this sRNA, which due to processing accumulates as three stabile 5' fragments in late exponential growth. Mobility shift assays using various mutants verified that theWe report on the characterization and target analysis of the small (s) RNA\(_{162}\) in the methanoarchaeon Methanosarcina mazei. Using a combination of genetic approaches, transcriptome analysis and computational predictions, the bicistronic MM2441-MM2440 mRNA encoding the transcription factor MM2441 and a protein of unknown function was identified as a potential target of this sRNA, which due to processing accumulates as three stabile 5' fragments in late exponential growth. Mobility shift assays using various mutants verified that the non-structured single-stranded linker region of sRNA\(_{162}\) (SLR) base-pairs with the MM2440-MM2441 mRNA internally, thereby masking the predicted ribosome binding site of MM2441. This most likely leads to translational repression of the second cistron resulting in dis-coordinated operon expression. Analysis of mutant RNAs in vivo confirmed that the SLR of sRNA\(_{162}\) is crucial for target interactions. Furthermore, our results indicate that sRNA\(_{162}\)-controlled MM2441 is involved in regulating the metabolic switch between the carbon sources methanol and methylamine. Moreover, biochemical studies demonstrated that the 50 end of sRNA\(_{162}\) targets the 5'-untranslated region of the cis-encoded MM2442 mRNA. Overall, this first study of archaeal sRNA/mRNA-target interactions unraveled that sRNA\(_{162}\) acts as an antisense (as) RNA on cis- and trans-encoded mRNAs via two distinct domains, indicating that cis-encoded asRNAs can have larger target regulons than previously anticipated.show moreshow less

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Metadaten
Author: Dominik Jäger, Sandy R. Pernitzsch, Andreas S. Richter, Rolf Backofen, Cynthia M. Sharma, Ruth A. Schmitz
URN:urn:nbn:de:bvb:20-opus-134972
Document Type:Journal article
Faculties:Medizinische Fakultät / Institut für Molekulare Infektionsbiologie
Language:English
Parent Title (English):Nucleic Acids Research
Year of Completion:2012
Volume:40
Issue:21
Pagenumber:10964-10979
Source:Nucleic Acids Research, 2012, Vol. 40, No. 21, p. 10964–10979. doi:10.1093/nar/gks847
DOI:https://doi.org/10.1093/nar/gks847
Dewey Decimal Classification:6 Technik, Medizin, angewandte Wissenschaften / 61 Medizin und Gesundheit / 610 Medizin und Gesundheit
Tag:GO1; acetivorans C2A; antisense RNAs; escherichia coli; methanol methyltransferase isozymes; methanosarcina mazei GO1; pyrococcus furiosus; small nucleolar RNAs; strain; transcriptional regulator; translational initiation
Release Date:2017/12/17
Licence (German):License LogoCC BY: Creative-Commons-Lizenz: Namensnennung