SRNA41 affects ribosome binding sites within polycistronic mRNAs in Methanosarcina mazei Gö1. Issue 5 (18th January 2018)

• Record Type: Journal Article
• Authors: Buddeweg, Anne; Sharma, Kundan; Urlaub, Henning; Schmitz, Ruth A.
• Is Part Of: Molecular microbiology. Volume 107:Issue 5(2018)
• Page Start: 595
• Page End: 609
• Publication Date: 2018-01-18
• Subjects: Molecular microbiology -- Periodicals; 572.829
• DOI: 10.1111/mmi.13900
• Languages: English
• ISSNs: 0950-382X

Abstract

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 sRNA41, highly expressed under N‐sufficiency. Comprising 120 nucleotides, sRNA41 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 sRNA41 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 sRNA41 ‐deletion mutant showed increased protein expression of both ACDS complexes in the absence of sRNA41, whereas no effect on transcript levels was detected, arguing for sRNA41 ‐mediated post‐transcriptional fine‐tuning of ACDS expression. We hypothesize that the physiological advantage of downregulating sRNA41 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 N2 ‐fixation, thus linking the carbon and N‐metabolism. Abstract : sRNA41 is a novel functionally characterized regulatory RNA in Methanosarcina mazei . It is the first archaeal sRNA capable to mask several ribosome binding sites within a polycistronic mRNA. Thereby sRNA41 inhibits translation initiation of the acetyl‐CoA decarbonylase/synthase (ACDS) multienzyme complex under nitrogen sufficient growth conditions. Under nitrogen limitation significantly decreased sRNA41 levels lead to higher amounts of ACDS, providing additional acetyl‐CoA required for synthesis of nitrogenase and accessory proteins.