Reactivity and Specificity of RNase T1, RNase A, and RNase H toward Oligonucleotides of RNA Containing 8-Oxo-7,8-dihydroguanosine.
Identifieur interne : 000836 ( Main/Exploration ); précédent : 000835; suivant : 000837Reactivity and Specificity of RNase T1, RNase A, and RNase H toward Oligonucleotides of RNA Containing 8-Oxo-7,8-dihydroguanosine.
Auteurs : Cassandra Herbert [États-Unis] ; Yannick Kokouvi Dzowo [États-Unis] ; Anthony Urban [États-Unis] ; Courtney N. Kiggins [États-Unis] ; Marino J E. Resendiz [États-Unis]Source :
- Biochemistry [ 1520-4995 ] ; 2018.
Descripteurs français
- KwdFr :
- ARN (), ARN (génétique), Dichroïsme circulaire, Guanosine (), Guanosine (analogues et dérivés), Guanosine (génétique), Humains, Oligonucléotides (), Oligonucléotides (génétique), Pancreatic ribonuclease (), Pancreatic ribonuclease (génétique), Ribonuclease H (), Ribonuclease H (génétique), Ribonuclease T1 (), Ribonuclease T1 (génétique), Ribonucléases (), Ribonucléases (génétique), Spécificité du substrat, Stress oxydatif (génétique).
- MESH :
- analogues et dérivés : Guanosine.
- génétique : ARN, Guanosine, Oligonucléotides, Pancreatic ribonuclease, Ribonuclease H, Ribonuclease T1, Ribonucléases, Stress oxydatif.
- ARN, Dichroïsme circulaire, Guanosine, Humains, Oligonucléotides, Pancreatic ribonuclease, Ribonuclease H, Ribonuclease T1, Ribonucléases, Spécificité du substrat.
English descriptors
- KwdEn :
- Circular Dichroism, Guanosine (analogs & derivatives), Guanosine (chemistry), Guanosine (genetics), Humans, Oligonucleotides (chemistry), Oligonucleotides (genetics), Oxidative Stress (genetics), RNA (chemistry), RNA (genetics), Ribonuclease H (chemistry), Ribonuclease H (genetics), Ribonuclease T1 (chemistry), Ribonuclease T1 (genetics), Ribonuclease, Pancreatic (chemistry), Ribonuclease, Pancreatic (genetics), Ribonucleases (chemistry), Ribonucleases (genetics), Substrate Specificity.
- MESH :
- chemical , analogs & derivatives : Guanosine.
- chemical , chemistry : Guanosine, Oligonucleotides, RNA, Ribonuclease H, Ribonuclease T1, Ribonuclease, Pancreatic, Ribonucleases.
- chemical , genetics : Guanosine, Oligonucleotides, RNA, Ribonuclease H, Ribonuclease T1, Ribonuclease, Pancreatic, Ribonucleases.
- genetics : Oxidative Stress.
- Circular Dichroism, Humans, Substrate Specificity.
Abstract
Understanding how oxidatively damaged RNA interacts with ribonucleases is important because of its proposed role in the development and progression of disease. Thus, understanding structural aspects of RNA containing lesions generated under oxidative stress, as well as its interactions with other biopolymers, is fundamental. We explored the reactivity of RNase A, RNase T1, and RNase H toward oligonucleotides of RNA containing 8-oxo-7,8-dihydroguanosine (8oxoG). This is the first example that addresses this relationship and will be useful for understanding (1) how these RNases can be used to characterize the structural impact that this lesion has on RNA and (2) how oxidatively modified RNA may be handled intracellularly. 8-OxoG was incorporated into 10-16-mers of RNA, and its reactivity with each ribonuclease was assessed via electrophoretic analyses, circular dichroism, and the use of other C8-purine-modified analogues (8-bromoguanosine, 8-methoxyguanosine, and 8-oxoadenosine). RNase T1 does not recognize sites containing 8-oxoG, while RNase A recognizes and cleaves RNA at positions containing this lesion while differentiating if it is involved in H-bonding. The selectivity of RNase A followed the order C > 8-oxoG ≈ U. In addition, isothermal titration calorimetry showed that an 8-oxoG-C3'-methylphosphate derivative can inhibit RNase A activity. Cleavage patterns obtained from RNase H displayed changes in reactivity in a sequence- and concentration-dependent manner and displayed recognition at sites containing the modification in some cases. These data will aid in understanding how this modification affects reactivity with ribonucleases and will enable the characterization of global and local structural changes in oxidatively damaged RNA.
DOI: 10.1021/acs.biochem.8b00277
PubMed: 29683663
Affiliations:
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Kiggins</name><affiliation wicri:level="1"><nlm:affiliation>Department of Chemistry , University of Colorado Denver , Science Building, 1151 Arapahoe Street , Denver , Colorado 80204 , United States.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Chemistry , University of Colorado Denver , Science Building, 1151 Arapahoe Street , Denver , Colorado 80204 </wicri:regionArea><wicri:noRegion>Colorado 80204 </wicri:noRegion></affiliation></author><author><name sortKey="Resendiz, Marino J E" sort="Resendiz, Marino J E" uniqKey="Resendiz M" first="Marino J E" last="Resendiz">Marino J E. Resendiz</name><affiliation wicri:level="1"><nlm:affiliation>Department of Chemistry , University of Colorado Denver , Science Building, 1151 Arapahoe Street , Denver , Colorado 80204 , United States.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Chemistry , University of Colorado Denver , Science Building, 1151 Arapahoe Street , Denver , Colorado 80204 </wicri:regionArea><wicri:noRegion>Colorado 80204 </wicri:noRegion></affiliation></author></analytic><series><title level="j">Biochemistry</title><idno type="eISSN">1520-4995</idno><imprint><date when="2018" type="published">2018</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Circular Dichroism</term><term>Guanosine (analogs & derivatives)</term><term>Guanosine (chemistry)</term><term>Guanosine (genetics)</term><term>Humans</term><term>Oligonucleotides (chemistry)</term><term>Oligonucleotides (genetics)</term><term>Oxidative Stress (genetics)</term><term>RNA (chemistry)</term><term>RNA (genetics)</term><term>Ribonuclease H (chemistry)</term><term>Ribonuclease H (genetics)</term><term>Ribonuclease T1 (chemistry)</term><term>Ribonuclease T1 (genetics)</term><term>Ribonuclease, Pancreatic (chemistry)</term><term>Ribonuclease, Pancreatic (genetics)</term><term>Ribonucleases (chemistry)</term><term>Ribonucleases (genetics)</term><term>Substrate Specificity</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>ARN ()</term><term>ARN (génétique)</term><term>Dichroïsme circulaire</term><term>Guanosine ()</term><term>Guanosine (analogues et dérivés)</term><term>Guanosine (génétique)</term><term>Humains</term><term>Oligonucléotides ()</term><term>Oligonucléotides (génétique)</term><term>Pancreatic ribonuclease ()</term><term>Pancreatic ribonuclease (génétique)</term><term>Ribonuclease H ()</term><term>Ribonuclease H (génétique)</term><term>Ribonuclease T1 ()</term><term>Ribonuclease T1 (génétique)</term><term>Ribonucléases ()</term><term>Ribonucléases (génétique)</term><term>Spécificité du substrat</term><term>Stress oxydatif (génétique)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="analogs & derivatives" xml:lang="en"><term>Guanosine</term></keywords><keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Guanosine</term><term>Oligonucleotides</term><term>RNA</term><term>Ribonuclease H</term><term>Ribonuclease T1</term><term>Ribonuclease, Pancreatic</term><term>Ribonucleases</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Guanosine</term><term>Oligonucleotides</term><term>RNA</term><term>Ribonuclease H</term><term>Ribonuclease T1</term><term>Ribonuclease, Pancreatic</term><term>Ribonucleases</term></keywords><keywords scheme="MESH" qualifier="analogues et dérivés" xml:lang="fr"><term>Guanosine</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Oxidative Stress</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>ARN</term><term>Guanosine</term><term>Oligonucléotides</term><term>Pancreatic ribonuclease</term><term>Ribonuclease H</term><term>Ribonuclease T1</term><term>Ribonucléases</term><term>Stress oxydatif</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Circular Dichroism</term><term>Humans</term><term>Substrate Specificity</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>ARN</term><term>Dichroïsme circulaire</term><term>Guanosine</term><term>Humains</term><term>Oligonucléotides</term><term>Pancreatic ribonuclease</term><term>Ribonuclease H</term><term>Ribonuclease T1</term><term>Ribonucléases</term><term>Spécificité du substrat</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Understanding how oxidatively damaged RNA interacts with ribonucleases is important because of its proposed role in the development and progression of disease. Thus, understanding structural aspects of RNA containing lesions generated under oxidative stress, as well as its interactions with other biopolymers, is fundamental. We explored the reactivity of RNase A, RNase T<sub>1</sub>, and RNase H toward oligonucleotides of RNA containing 8-oxo-7,8-dihydroguanosine (8oxoG). This is the first example that addresses this relationship and will be useful for understanding (1) how these RNases can be used to characterize the structural impact that this lesion has on RNA and (2) how oxidatively modified RNA may be handled intracellularly. 8-OxoG was incorporated into 10-16-mers of RNA, and its reactivity with each ribonuclease was assessed via electrophoretic analyses, circular dichroism, and the use of other C8-purine-modified analogues (8-bromoguanosine, 8-methoxyguanosine, and 8-oxoadenosine). RNase T<sub>1</sub> does not recognize sites containing 8-oxoG, while RNase A recognizes and cleaves RNA at positions containing this lesion while differentiating if it is involved in H-bonding. The selectivity of RNase A followed the order C > 8-oxoG ≈ U. In addition, isothermal titration calorimetry showed that an 8-oxoG-C3'-methylphosphate derivative can inhibit RNase A activity. Cleavage patterns obtained from RNase H displayed changes in reactivity in a sequence- and concentration-dependent manner and displayed recognition at sites containing the modification in some cases. These data will aid in understanding how this modification affects reactivity with ribonucleases and will enable the characterization of global and local structural changes in oxidatively damaged RNA.</div></front></TEI><affiliations><list><country><li>États-Unis</li></country></list><tree><country name="États-Unis"><noRegion><name sortKey="Herbert, Cassandra" sort="Herbert, Cassandra" uniqKey="Herbert C" first="Cassandra" last="Herbert">Cassandra Herbert</name></noRegion><name sortKey="Dzowo, Yannick Kokouvi" sort="Dzowo, Yannick Kokouvi" uniqKey="Dzowo Y" first="Yannick Kokouvi" last="Dzowo">Yannick Kokouvi Dzowo</name><name sortKey="Kiggins, Courtney N" sort="Kiggins, Courtney N" uniqKey="Kiggins C" first="Courtney N" last="Kiggins">Courtney N. Kiggins</name><name sortKey="Resendiz, Marino J E" sort="Resendiz, Marino J E" uniqKey="Resendiz M" first="Marino J E" last="Resendiz">Marino J E. Resendiz</name><name sortKey="Urban, Anthony" sort="Urban, Anthony" uniqKey="Urban A" first="Anthony" last="Urban">Anthony Urban</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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