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sRNA 41 affects ribosome binding sites within polycistronic mRNAs in Methanosarcina mazei Gö1
Author(s) -
Buddeweg Anne,
Sharma Kundan,
Urlaub Henning,
Schmitz Ruth A.
Publication year - 2018
Publication title -
molecular microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.857
H-Index - 247
eISSN - 1365-2958
pISSN - 0950-382X
DOI - 10.1111/mmi.13900
Subject(s) - biology , methanosarcina , ribosome , messenger rna , transfer rna , genetics , microbiology and biotechnology , rna , gene , archaea
Summary Several noncoding RNAs potentially involved in nitrogen (N)‐regulation have been detected in Methanosarcina mazei , however, targets have been identified only for one of them. Here, we report on the function of sRNA 41 , highly expressed under N‐sufficiency. Comprising 120 nucleotides, sRNA 41 shows high sequence and structural conservation within draft genomes of numerous Methanosarcina species. In silico target prediction revealed several potential targets, including genes of two homologous operons encoding for acetyl‐CoA‐decarbonylase/synthase complexes (ACDS) representing highly probable target candidates. A highly conserved single stranded region of sRNA 41 was predicted to mask six independent ribosome binding sites of these two polycistronic mRNAs and was verified in vitro by microscale thermophoresis. Proteome analysis of the respective sRNA 41 ‐deletion mutant showed increased protein expression of both ACDS complexes in the absence of sRNA 41 , whereas no effect on transcript levels was detected, arguing for sRNA 41 ‐mediated post‐transcriptional fine‐tuning of ACDS expression. We hypothesize that the physiological advantage of downregulating sRNA 41 under N‐limiting conditions is the resulting increase of ACDS protein levels. This provides sufficient amounts of amino acids for nitrogenase synthesis as well as reducing equivalents and energy for N 2 ‐fixation, thus linking the carbon and N‐metabolism.