Genome-wide mRNA processing in methanogenic archaea reveals post-transcriptional regulation of ribosomal protein synthesis

Nucleic Acids Res. 2017 Jul 7;45(12):7285-7298. doi: 10.1093/nar/gkx454.

Abstract

Unlike stable RNAs that require processing for maturation, prokaryotic cellular mRNAs generally follow an 'all-or-none' pattern. Herein, we used a 5΄ monophosphate transcript sequencing (5΄P-seq) that specifically captured the 5΄-end of processed transcripts and mapped the genome-wide RNA processing sites (PSSs) in a methanogenic archaeon. Following statistical analysis and stringent filtration, we identified 1429 PSSs, among which 23.5% and 5.4% were located in 5΄ untranslated region (uPSS) and intergenic region (iPSS), respectively. A predominant uridine downstream PSSs served as a processing signature. Remarkably, 5΄P-seq detected overrepresented uPSS and iPSS in the polycistronic operons encoding ribosomal proteins, and the majority upstream and proximal ribosome binding sites, suggesting a regulatory role of processing on translation initiation. The processed transcripts showed increased stability and translation efficiency. Particularly, processing within the tricistronic transcript of rplA-rplJ-rplL enhanced the translation of rplL, which can provide a driving force for the 1:4 stoichiometry of L10 to L12 in the ribosome. Growth-associated mRNA processing intensities were also correlated with the cellular ribosomal protein levels, thereby suggesting that mRNA processing is involved in tuning growth-dependent ribosome synthesis. In conclusion, our findings suggest that mRNA processing-mediated post-transcriptional regulation is a potential mechanism of ribosomal protein synthesis and stoichiometry.

MeSH terms

  • Archaeal Proteins / genetics*
  • Archaeal Proteins / metabolism
  • Base Sequence
  • Cloning, Molecular
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Gene Expression
  • Genome, Archaeal*
  • Methanococcus / genetics*
  • Methanococcus / metabolism
  • Methanosarcinaceae / genetics*
  • Methanosarcinaceae / metabolism
  • Nucleic Acid Conformation
  • Peptide Chain Initiation, Translational
  • Protein Isoforms / genetics
  • Protein Isoforms / metabolism
  • RNA Processing, Post-Transcriptional*
  • RNA, Messenger / chemistry
  • RNA, Messenger / genetics*
  • RNA, Messenger / metabolism
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Ribosomal Proteins / genetics*
  • Ribosomal Proteins / metabolism

Substances

  • Archaeal Proteins
  • Protein Isoforms
  • RNA, Messenger
  • Recombinant Proteins
  • Ribosomal Proteins
  • ribosomal protein L7-L12