A tertiary structural element in S box leader RNAs is required for S-adenosylmethionine-directed transcription termination.

Abstract:

:The S box transcription termination control system regulates expression of genes involved in methionine metabolism. Expression of the S box regulon, comprised of 11 transcriptional units in Bacillus subtilis, is induced in response to starvation for methionine. We previously demonstrated that S-adenosylmethionine (SAM) is the molecular effector sensed by the S box leader RNAs during transcription. A secondary structure model for S box leader RNAs was developed based on conservation of primary sequence elements and sequence covariation in helical domains. Covariation of nucleotides in two distantly spaced unpaired regions in the S box leader RNAs suggested that these two domains might interact in the RNA tertiary structure. In this study, site-directed mutagenesis of the covarying residues in two B. subtilis S box leader sequences was employed to test the hypothesis that base-pairing between these regions may be important. The effect of these mutations on in vivo expression, transcription termination in vitro, SAM binding, and leader RNA structure strongly supported the model that interaction between these two regions plays a key role in S box leader function. This provides the first insight into the three-dimensional arrangement of structural elements within the S box RNAs.

journal_name

Mol Microbiol

journal_title

Molecular microbiology

authors

McDaniel BA,Grundy FJ,Henkin TM

doi

10.1111/j.1365-2958.2005.04740.x

subject

Has Abstract

pub_date

2005-08-01 00:00:00

pages

1008-21

issue

4

eissn

0950-382X

issn

1365-2958

pii

MMI4740

journal_volume

57

pub_type

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