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Mapping interactions between the RNA chaperone FinO and its RNA targets

Bacterial conjugation is regulated by two-component repression comprising the antisense RNA FinP, and its protein co-factor FinO. FinO mediates base-pairing of FinP to the 5′-untranslated region (UTR) of traJ mRNA, which leads to translational inhibition of the transcriptional activator TraJ and sub...

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Autores principales: Arthur, David C., Edwards, Ross A., Tsutakawa, Susan, Tainer, John A., Frost, Laura S., Glover, J. N. Mark
Formato: Texto
Lenguaje:English
Publicado: Oxford University Press 2011
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3105414/
https://www.ncbi.nlm.nih.gov/pubmed/21278162
http://dx.doi.org/10.1093/nar/gkr025
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author Arthur, David C.
Edwards, Ross A.
Tsutakawa, Susan
Tainer, John A.
Frost, Laura S.
Glover, J. N. Mark
author_facet Arthur, David C.
Edwards, Ross A.
Tsutakawa, Susan
Tainer, John A.
Frost, Laura S.
Glover, J. N. Mark
author_sort Arthur, David C.
collection PubMed
description Bacterial conjugation is regulated by two-component repression comprising the antisense RNA FinP, and its protein co-factor FinO. FinO mediates base-pairing of FinP to the 5′-untranslated region (UTR) of traJ mRNA, which leads to translational inhibition of the transcriptional activator TraJ and subsequent down regulation of conjugation genes. Yet, little is known about how FinO binds to its RNA targets or how this interaction facilitates FinP and traJ mRNA pairing. Here, we use solution methods to determine how FinO binds specifically to its minimal high affinity target, FinP stem–loop II (SLII), and its complement SLIIc from traJ mRNA. Ribonuclease footprinting reveals that FinO contacts the base of the stem and the 3′ single-stranded tails of these RNAs. The phosphorylation or oxidation of the 3′-nucleotide blocks FinO binding, suggesting FinO binds the 3′-hydroxyl of its RNA targets. The collective results allow the generation of an energy-minimized model of the FinO–SLII complex, consistent with small-angle X-ray scattering data. The repression complex model was constrained using previously reported cross-linking data and newly developed footprinting results. Together, these data lead us to propose a model of how FinO mediates FinP/traJ mRNA pairing to down regulate bacterial conjugation.
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spelling pubmed-31054142011-06-01 Mapping interactions between the RNA chaperone FinO and its RNA targets Arthur, David C. Edwards, Ross A. Tsutakawa, Susan Tainer, John A. Frost, Laura S. Glover, J. N. Mark Nucleic Acids Res Structural Biology Bacterial conjugation is regulated by two-component repression comprising the antisense RNA FinP, and its protein co-factor FinO. FinO mediates base-pairing of FinP to the 5′-untranslated region (UTR) of traJ mRNA, which leads to translational inhibition of the transcriptional activator TraJ and subsequent down regulation of conjugation genes. Yet, little is known about how FinO binds to its RNA targets or how this interaction facilitates FinP and traJ mRNA pairing. Here, we use solution methods to determine how FinO binds specifically to its minimal high affinity target, FinP stem–loop II (SLII), and its complement SLIIc from traJ mRNA. Ribonuclease footprinting reveals that FinO contacts the base of the stem and the 3′ single-stranded tails of these RNAs. The phosphorylation or oxidation of the 3′-nucleotide blocks FinO binding, suggesting FinO binds the 3′-hydroxyl of its RNA targets. The collective results allow the generation of an energy-minimized model of the FinO–SLII complex, consistent with small-angle X-ray scattering data. The repression complex model was constrained using previously reported cross-linking data and newly developed footprinting results. Together, these data lead us to propose a model of how FinO mediates FinP/traJ mRNA pairing to down regulate bacterial conjugation. Oxford University Press 2011-05 2011-01-27 /pmc/articles/PMC3105414/ /pubmed/21278162 http://dx.doi.org/10.1093/nar/gkr025 Text en © The Author(s) 2011. Published by Oxford University Press. http://creativecommons.org/licenses/by-nc/2.5 This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Structural Biology
Arthur, David C.
Edwards, Ross A.
Tsutakawa, Susan
Tainer, John A.
Frost, Laura S.
Glover, J. N. Mark
Mapping interactions between the RNA chaperone FinO and its RNA targets
title Mapping interactions between the RNA chaperone FinO and its RNA targets
title_full Mapping interactions between the RNA chaperone FinO and its RNA targets
title_fullStr Mapping interactions between the RNA chaperone FinO and its RNA targets
title_full_unstemmed Mapping interactions between the RNA chaperone FinO and its RNA targets
title_short Mapping interactions between the RNA chaperone FinO and its RNA targets
title_sort mapping interactions between the rna chaperone fino and its rna targets
topic Structural Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3105414/
https://www.ncbi.nlm.nih.gov/pubmed/21278162
http://dx.doi.org/10.1093/nar/gkr025
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