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Ribonuclease D Processes a Small RNA Regulator of Multicellular Development in Myxobacteria.

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Ribonuclease D (RNase D) is crucial for processing the small RNA Pxr, which controls bacterial development in Myxococcus xanthus. Disrupting RNase D prevents Pxr maturation, halting development and revealing its role in sRNA processing.

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Area of Science:

  • Bacterial genetics and molecular biology
  • Microbial development and signaling
  • RNA processing and regulation

Background:

  • Non-coding small RNAs (sRNAs) regulate gene expression in bacteria.
  • In *Myxococcus xanthus*, the sRNA Pxr controls the transition from vegetative growth to multicellular development.
  • Pxr inhibits development under nutrient-rich conditions, with inhibition relieved upon starvation.

Purpose of the Study:

  • To identify genes essential for the function of the sRNA Pxr in *Myxococcus xanthus* development.
  • To elucidate the role of Ribonuclease D (RNase D) in Pxr maturation and function.
  • To understand the mechanism of Pxr processing and its impact on bacterial life-cycle transitions.

Main Methods:

  • Transposon mutagenesis of a developmentally defective *Myxococcus xanthus* strain (OC) to identify suppressor mutations.
  • Analysis of Pxr processing and accumulation in wild-type and mutant strains.
  • In vitro processing assays using purified RNase D and Pxr transcripts.

Main Results:

  • Disruption of the *rnd* gene, encoding RNase D, abolished the accumulation of the active sRNA Pxr-S.
  • Loss of RNase D led to increased accumulation of a longer Pxr precursor transcript (Pxr-XL) and decreased Pxr-S.
  • RNase D was shown to process Pxr-XL into Pxr-L in vitro, indicating a two-step processing pathway for Pxr maturation.

Conclusions:

  • Housekeeping ribonuclease RNase D plays a central role in the maturation of the developmental regulatory sRNA Pxr.
  • Pxr sRNA maturation involves a sequential two-step processing pathway, with RNase D acting in the initial step.
  • This study provides the first evidence implicating RNase D in sRNA processing, highlighting its importance beyond tRNA maturation.