Abstract
6S RNA, a conserved and abundant small non-coding RNA
found in most bacteria, regulates gene expression by inhibiting
RNA polymerase (RNAP) holoenzyme. 6S RNAs from
α-proteobacteria have been studied poorly so far. Here, we
present a first in-depth analysis of 6S RNAs from two α-proteobacteria
species, Bradyrhizobium japonicum and Sinorhizobium
meliloti. Although both belong to the order Rhizobiales
and are typical nitrogen-fixing symbionts of legumes,
their 6S RNA expression profiles were found to differ: B. japonicum
6S RNA accumulated in the stationary phase, thus
being reminiscent of Escherichia coli 6S RNA, whereas S. meliloti
6S RNA level peaked at the transition to the stationary
phase, similarly to Rhodobacter sphaeroides 6S RNA. We demonstrated
in vitro that both RNAs have hallmarks of 6S
RNAs: they bind to the σ70-type RNAP holoenzyme and serve
as templates for de novo transcription of so-called product
RNAs (pRNAs) ranging in length from ~13 to 24 nucleotides,
with further evidence of the synthesis of even longer pRNAs.
Likewise, stably bound pRNAs were found to rearrange the
6S RNA structure to induce its dissociation from RNAP.
Compared with B. japonicum 6S RNA, considerable conformational
heterogeneity was observed for S. meliloti 6S RNA
and its complexes with pRNAs, even though the two 6S RNAs
share ~75% sequence identity. Overall, our findings suggest
that the two rhizobial 6S RNAs have diverged with respect to
their regulatory impact on gene expression throughout the
bacterial life cycle.
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