Poster Presentation 51st Lorne Proteins Conference 2026

sRNAs, The Jack of All Trades in Bacteria Optimizing Growth and Minimizing Energy Costs Through Post-transcriptional Regulation. (#405)

Kwok Jian Goh 1 2 , Sophie Wharrie 1 3 4 , Hee-Jung Shim 1 3 4 , Trevor Lithgow 1 2
  1. ARC Centre of Excellence for the Mathematical Analysis of Cellular Systems (MACSYS), Monash University, Clayton, Victoria, Australia
  2. Department of Microbiology, Monash Biomedicine Discovery Institute, Monash University, Clayton, Victoria, Australia
  3. Melbourne Integrative Genomics, The University of Melbourne, Parkville, Victoria, Australia
  4. School of Mathematics and Statistics, Faculty of Science, The University of Melbourne, Parkville, Victoria, Australia

The central dogma is now understood to be extensively modulated by regulatory mechanisms acting before and after both transcription and translation. Small RNAs (sRNAs) are generally non‑coding RNAs that regulate the translation and stability of mRNAs in prokaryotes. The sRNAs typically base‑pair with target mRNAs with limited complementarity, often with the assistance of the RNA‑binding protein Hfq, and frequently act near the ribosome‑binding site to inhibit translation initiation or within the open reading frame to promote translational arrest and mRNA decay. To globally identify RNA–RNA interactions in pathogenic Klebsiella, RIL‑seq was applied to the hypervirulent K1 capsule type clinical strain SGH10 in our previous work [1]. This high‑throughput approach yielded numerous RNA–RNA contacts, from which we focused on interactions associated with membrane biogenesis. A subset of these was prioritized as putative bona fide sRNA–mRNA regulatory pairs, and their activity was interrogated using GFP reporter assays. We identified a novel regulatory interaction in which elevated levels of the sRNA SdsR repress translation of TamA, the outer membrane component of the translocation and assembly module (TAM). Computational predictions indicated that SdsR–tamA base‑pairing occurs in the vicinity of the tamA start codon. Guided by this, we engineered an SdsR mutant that abolished repression of tamA in reporter assays. However, a compensatory mutation introduced into tamA only partially restored SdsR‑dependent repression, suggesting additional structural or contextual features contribute to this interaction. Using antibodies against TamA as well as TolC, a known SdsR target, we were able to assess the impact of the SdsR mutant with their abundances detected on Western blot as the output. Together, these findings expand the current understanding of sRNA‑mediated control of envelope‑associated functions in Klebsiella and provide a foundation for future integration of sRNA networks into whole‑cell predictive models.

  1. Goh, K.J., Altuvia, Y., Argaman, L., Raz, Y., Bar, A., Lithgow, T., Margalit, H. and Gan, Y.H., 2024. RIL-seq reveals extensive involvement of small RNAs in virulence and capsule regulation in hypervirulent Klebsiella pneumoniae. Nucleic Acids Research, 52(15), pp.9119-9138.