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Positive regulation by small RNAs and the role of Hfq.
Proc Natl Acad Sci U S A. 2010 May 25; 107(21):9602-7.PN

Abstract

Bacterial small noncoding RNAs carry out both positive and negative regulation of gene expression by pairing with mRNAs; in Escherichia coli, this regulation often requires the RNA chaperone Hfq. Three small regulatory RNAs (sRNAs), DsrA, RprA, and ArcZ, positively regulate translation of the sigma factor RpoS, each pairing with the 5' leader to open up an inhibitory hairpin. In vitro, rpoS interaction with sRNAs depends upon an (AAN)(4) Hfq-binding site upstream of the pairing region. Here we show that both Hfq and this Hfq binding site are required for RprA or ArcZ to act in vivo and to form a stable complex with rpoS mRNA in vitro; both were partially dispensable for DsrA at 37 degrees C. ArcZ sRNA is processed from 121 nt to a stable 56 nt species that contains the pairing region; only the 56 nt ArcZ makes a strong Hfq-dependent complex with rpoS. For each of these sRNAs, the stability of the sRNA*mRNA complexes, rather than their rate of formation, best predicted in vivo activity. These studies demonstrate that binding of Hfq to the rpoS mRNA is critical for sRNA regulation under normal conditions, but if the stability of the sRNA*mRNA complex is sufficiently high, the requirement for Hfq can be bypassed.

Authors+Show Affiliations

Johns Hopkins University, Program in Cell, Molecular, Developmental Biology and Biophysics, Baltimore, MD 21218, USA.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, N.I.H., Extramural
Research Support, N.I.H., Intramural

Language

eng

PubMed ID

20457943

Citation

Soper, Toby, et al. "Positive Regulation By Small RNAs and the Role of Hfq." Proceedings of the National Academy of Sciences of the United States of America, vol. 107, no. 21, 2010, pp. 9602-7.
Soper T, Mandin P, Majdalani N, et al. Positive regulation by small RNAs and the role of Hfq. Proc Natl Acad Sci U S A. 2010;107(21):9602-7.
Soper, T., Mandin, P., Majdalani, N., Gottesman, S., & Woodson, S. A. (2010). Positive regulation by small RNAs and the role of Hfq. Proceedings of the National Academy of Sciences of the United States of America, 107(21), 9602-7. https://doi.org/10.1073/pnas.1004435107
Soper T, et al. Positive Regulation By Small RNAs and the Role of Hfq. Proc Natl Acad Sci U S A. 2010 May 25;107(21):9602-7. PubMed PMID: 20457943.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Positive regulation by small RNAs and the role of Hfq. AU - Soper,Toby, AU - Mandin,Pierre, AU - Majdalani,Nadim, AU - Gottesman,Susan, AU - Woodson,Sarah A, Y1 - 2010/05/10/ PY - 2010/5/12/entrez PY - 2010/5/12/pubmed PY - 2010/6/30/medline SP - 9602 EP - 7 JF - Proceedings of the National Academy of Sciences of the United States of America JO - Proc Natl Acad Sci U S A VL - 107 IS - 21 N2 - Bacterial small noncoding RNAs carry out both positive and negative regulation of gene expression by pairing with mRNAs; in Escherichia coli, this regulation often requires the RNA chaperone Hfq. Three small regulatory RNAs (sRNAs), DsrA, RprA, and ArcZ, positively regulate translation of the sigma factor RpoS, each pairing with the 5' leader to open up an inhibitory hairpin. In vitro, rpoS interaction with sRNAs depends upon an (AAN)(4) Hfq-binding site upstream of the pairing region. Here we show that both Hfq and this Hfq binding site are required for RprA or ArcZ to act in vivo and to form a stable complex with rpoS mRNA in vitro; both were partially dispensable for DsrA at 37 degrees C. ArcZ sRNA is processed from 121 nt to a stable 56 nt species that contains the pairing region; only the 56 nt ArcZ makes a strong Hfq-dependent complex with rpoS. For each of these sRNAs, the stability of the sRNA*mRNA complexes, rather than their rate of formation, best predicted in vivo activity. These studies demonstrate that binding of Hfq to the rpoS mRNA is critical for sRNA regulation under normal conditions, but if the stability of the sRNA*mRNA complex is sufficiently high, the requirement for Hfq can be bypassed. SN - 1091-6490 UR - https://www.unboundmedicine.com/medline/citation/20457943/Positive_regulation_by_small_RNAs_and_the_role_of_Hfq_ L2 - http://www.pnas.org/cgi/pmidlookup?view=long&pmid=20457943 DB - PRIME DP - Unbound Medicine ER -