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Small non-coding RNAs (ncRNAs) are a class of regulatory molecules, which remain understudied in bacteria. In the extremophilic bacterium  Deinococcus radiodurans , although hundreds of ncRNAs have been identified, few have been characterized in detail. In this study, we report the identification and characterization of a novel heat-inducible ncRNA named  DnrH . Heat tolerance analysis showed that deleting  DnrH  significantly inhibited viability in response to high temperature conditions. Comparative phenotypic and qRT-PCR analyses of a  DnrH  mutant (Δ DnrH ) and wild-type (WT)  D. radiodurans  suggested that  DnrH  is potentially involved in regulating the expression of the heat shock-related gene  Hsp20 . Microscale thermophoresis and genetic complementation showed that a 28-nucleotide (nt) sequence in the stem-loop structure of  DnrH  (143–170 nt) pairs with its counterpart in the coding region of  Hsp20  mRNA (91–117 nt) via a 22 nt region.  In vivo , mutation of the 22-nt region in the  D. radiodurans  genome led to a reduction in heat tolerance similar to that observed in the  DnrH -mutant. Our results show that  DnrH  positively influences heat tolerance by increasing the transcription of  Hsp20  mRNA, demonstrating, for the first time, a ncRNA that directly controls the expression of a heat stress-resistance gene. This work provides new insight into the heat stress response mechanism of  D. radiodurans  as well as other extremophiles that express similar Hsp20 proteins.

Targeting Hsp20 Using the Novel Small Non-coding RNA DnrH Regulates Heat Tolerance in Deinococcus radiodurans