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Effect of Hfq on RprA-rpoS mRNA pairing: Hfq-RNA binding and the influence of the 5' rpoS mRNA leader region.
Biochemistry. 2008 Oct 28; 47(43):11184-95.B

Abstract

The rpoS mRNA encodes a stress response transcription factor in Escherichia coli. It is one of a growing number of mRNAs found to be regulated by small RNAs (sRNA). Translation initiation of rpoS mRNA is enhanced by two sRNAs, DsrA and RprA, that pair to the same site near the rpoS start codon in the presence of the Hfq protein. In this work, we examine the interaction of E. coli Hfq with RprA and two portions of the rpoS mRNA leader region. One rpoS RNA, rpoS-L, contained the entire 565-nucleotide leader region, while the other, rpoS-S, contained the 199-nucleotide sequence surrounding the start codon. An RNase H assay indicated both rpoS RNAs have similar secondary structures in the translation initiation region. Hfq formed two complexes with RprA in a gel mobility assay with binding parameters similar to values previously determined for DsrA. Unlike DsrA, Hfq binding to RprA was inhibited by poly(A) and influenced by Hfq mutations on both the distal and proximal surfaces. Hfq increased the level of RprA binding to both rpoS RNAs but showed a much larger enhancement when rpoS-L, the entire leader region, was examined. The lower affinity of RprA for rpoS-L versus rpoS-S in the absence of Hfq suggests that Hfq overcomes an inhibitory structure within rpoS-L in stimulating RprA binding. Similar results were obtained with DsrA. The results indicate that the full upstream leader sequence of rpoS mRNA influences Hfq-facilitated annealing of RprA and DsrA and is likely to be involved in its regulation.

Authors+Show Affiliations

School of Biology and Institute of Bioengineering and Biosciences, Georgia Institute of Technology, Atlanta, Georgia 30332, USA.No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

18826256

Citation

Updegrove, Taylor, et al. "Effect of Hfq On RprA-rpoS mRNA Pairing: Hfq-RNA Binding and the Influence of the 5' rpoS mRNA Leader Region." Biochemistry, vol. 47, no. 43, 2008, pp. 11184-95.
Updegrove T, Wilf N, Sun X, et al. Effect of Hfq on RprA-rpoS mRNA pairing: Hfq-RNA binding and the influence of the 5' rpoS mRNA leader region. Biochemistry. 2008;47(43):11184-95.
Updegrove, T., Wilf, N., Sun, X., & Wartell, R. M. (2008). Effect of Hfq on RprA-rpoS mRNA pairing: Hfq-RNA binding and the influence of the 5' rpoS mRNA leader region. Biochemistry, 47(43), 11184-95. https://doi.org/10.1021/bi800479p
Updegrove T, et al. Effect of Hfq On RprA-rpoS mRNA Pairing: Hfq-RNA Binding and the Influence of the 5' rpoS mRNA Leader Region. Biochemistry. 2008 Oct 28;47(43):11184-95. PubMed PMID: 18826256.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Effect of Hfq on RprA-rpoS mRNA pairing: Hfq-RNA binding and the influence of the 5' rpoS mRNA leader region. AU - Updegrove,Taylor, AU - Wilf,Nabil, AU - Sun,Xueguang, AU - Wartell,Roger M, Y1 - 2008/10/01/ PY - 2008/10/2/pubmed PY - 2008/11/15/medline PY - 2008/10/2/entrez SP - 11184 EP - 95 JF - Biochemistry JO - Biochemistry VL - 47 IS - 43 N2 - The rpoS mRNA encodes a stress response transcription factor in Escherichia coli. It is one of a growing number of mRNAs found to be regulated by small RNAs (sRNA). Translation initiation of rpoS mRNA is enhanced by two sRNAs, DsrA and RprA, that pair to the same site near the rpoS start codon in the presence of the Hfq protein. In this work, we examine the interaction of E. coli Hfq with RprA and two portions of the rpoS mRNA leader region. One rpoS RNA, rpoS-L, contained the entire 565-nucleotide leader region, while the other, rpoS-S, contained the 199-nucleotide sequence surrounding the start codon. An RNase H assay indicated both rpoS RNAs have similar secondary structures in the translation initiation region. Hfq formed two complexes with RprA in a gel mobility assay with binding parameters similar to values previously determined for DsrA. Unlike DsrA, Hfq binding to RprA was inhibited by poly(A) and influenced by Hfq mutations on both the distal and proximal surfaces. Hfq increased the level of RprA binding to both rpoS RNAs but showed a much larger enhancement when rpoS-L, the entire leader region, was examined. The lower affinity of RprA for rpoS-L versus rpoS-S in the absence of Hfq suggests that Hfq overcomes an inhibitory structure within rpoS-L in stimulating RprA binding. Similar results were obtained with DsrA. The results indicate that the full upstream leader sequence of rpoS mRNA influences Hfq-facilitated annealing of RprA and DsrA and is likely to be involved in its regulation. SN - 1520-4995 UR - https://www.unboundmedicine.com/medline/citation/18826256/Effect_of_Hfq_on_RprA_rpoS_mRNA_pairing:_Hfq_RNA_binding_and_the_influence_of_the_5'_rpoS_mRNA_leader_region_ L2 - https://doi.org/10.1021/bi800479p DB - PRIME DP - Unbound Medicine ER -