Relationship between 2'-hydroxyls and magnesium binding in the hammerhead RNA domain: a model for ribozyme catalysis
The use of deoxyribonucleotide substitution in RNA (mixed RNA/DNA polymers) permits an evaluation of the role of 2'-hydroxyl groups in ribozyme catalysis. Specific deoxyribonucleotide substitution at G9 and A13 of the ribozyme decreases the catalytic activity (kcat) of the ribozyme by factors o...
Gespeichert in:
| Veröffentlicht in: | Biochemistry (Easton) 1991-04, Vol.30 (16), p.4020-4025 |
|---|---|
| Hauptverfasser: | , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 4025 |
|---|---|
| container_issue | 16 |
| container_start_page | 4020 |
| container_title | Biochemistry (Easton) |
| container_volume | 30 |
| creator | Perreault, Jean Pierre Labuda, Damian Usman, Nassim Yang, Jing Hua Cedergren, Robert |
| description | The use of deoxyribonucleotide substitution in RNA (mixed RNA/DNA polymers) permits an evaluation of the role of 2'-hydroxyl groups in ribozyme catalysis. Specific deoxyribonucleotide substitution at G9 and A13 of the ribozyme decreases the catalytic activity (kcat) of the ribozyme by factors of 14 and 20, respectively. The reduction of the reaction rate concomitant with the absence of these 2'-OHs or the 2'-OH of the substrate U7 position can be partially compensated by increasing the Mg2+ concentration above 10 mM. The KMg of the all-RNA ribozyme is 5.3 mM, and the lack of either of the three influential 2'-OHs increases this value by a factor of approximately 3. These and other reaction constants for the ribozyme and the deoxy-substituted analogues have been determined by assuming a three-step mechanism. The data presented here provide the basis for the formulation of a molecular model of ribozyme activity. |
| doi_str_mv | 10.1021/bi00230a029 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_80525458</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>80525458</sourcerecordid><originalsourceid>FETCH-LOGICAL-a481t-562d73f0d7dde829f3980a7ccdb9765a504c81a20b781188122526888bfb2d7e3</originalsourceid><addsrcrecordid>eNqF0TFv1TAQB3ALgcprYWJG8gIdUODsxLHTrapoQaoAlcJqXWKnzyW2H3YiGj49QXmiDEhMlvX_-eS7I-QZg9cMOHvTOgBeAgJvHpANExyKqmnEQ7IBgLrgTQ2PyWHOt8u1AlkdkAMmQXElNmS8sgOOLoa8dTva2vGHtYHy42I7mxTv5iFTDIZ6vAk2u8nT1gXjwg11gY5bS7fovU1bi4ZefTilJnp04YQi9dHYgfYx0eTa-HP2lnY44jBnl5-QRz0O2T7dn0fky_nb67N3xeXHi_dnp5cFVoqNhai5kWUPRhpjFW_6slGAsutM28haoICqUww5tFIxphTjXPBaKdX27fLSlkfk5Vp3l-L3yeZRe5c7OwwYbJyyViC4qIT6L2SiqWqm5AJfrbBLMedke71LzmOaNQP9exn6r2Us-vm-7NR6a-7tOv0lf7HPMXc49AlD5_I9a-TSIrDFFatzebR3f3JM33QtSyn09afP-kKwkn0tha4Xf7x67LK-jVMKy5T_-cNf6WqsHA</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>15946187</pqid></control><display><type>article</type><title>Relationship between 2'-hydroxyls and magnesium binding in the hammerhead RNA domain: a model for ribozyme catalysis</title><source>MEDLINE</source><source>ACS Publications</source><creator>Perreault, Jean Pierre ; Labuda, Damian ; Usman, Nassim ; Yang, Jing Hua ; Cedergren, Robert</creator><creatorcontrib>Perreault, Jean Pierre ; Labuda, Damian ; Usman, Nassim ; Yang, Jing Hua ; Cedergren, Robert</creatorcontrib><description>The use of deoxyribonucleotide substitution in RNA (mixed RNA/DNA polymers) permits an evaluation of the role of 2'-hydroxyl groups in ribozyme catalysis. Specific deoxyribonucleotide substitution at G9 and A13 of the ribozyme decreases the catalytic activity (kcat) of the ribozyme by factors of 14 and 20, respectively. The reduction of the reaction rate concomitant with the absence of these 2'-OHs or the 2'-OH of the substrate U7 position can be partially compensated by increasing the Mg2+ concentration above 10 mM. The KMg of the all-RNA ribozyme is 5.3 mM, and the lack of either of the three influential 2'-OHs increases this value by a factor of approximately 3. These and other reaction constants for the ribozyme and the deoxy-substituted analogues have been determined by assuming a three-step mechanism. The data presented here provide the basis for the formulation of a molecular model of ribozyme activity.</description><identifier>ISSN: 0006-2960</identifier><identifier>EISSN: 1520-4995</identifier><identifier>DOI: 10.1021/bi00230a029</identifier><identifier>PMID: 1708285</identifier><language>eng</language><publisher>Washington, DC: American Chemical Society</publisher><subject>Base Sequence ; Binding Sites ; Biological and medical sciences ; Catalysis ; DNA - chemical synthesis ; Fundamental and applied biological sciences. Psychology ; Hydroxylation ; Kinetics ; Magnesium - chemistry ; Mechanisms. Catalysis. Electron transfer. Models ; Models, Genetic ; Molecular biophysics ; Molecular Sequence Data ; Nucleic Acid Conformation ; Physical chemistry in biology ; RNA - chemical synthesis ; RNA - chemistry ; RNA, Catalytic - metabolism ; Thermodynamics</subject><ispartof>Biochemistry (Easton), 1991-04, Vol.30 (16), p.4020-4025</ispartof><rights>1991 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-a481t-562d73f0d7dde829f3980a7ccdb9765a504c81a20b781188122526888bfb2d7e3</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://pubs.acs.org/doi/pdf/10.1021/bi00230a029$$EPDF$$P50$$Gacs$$H</linktopdf><linktohtml>$$Uhttps://pubs.acs.org/doi/10.1021/bi00230a029$$EHTML$$P50$$Gacs$$H</linktohtml><link.rule.ids>314,780,784,2764,27075,27923,27924,56737,56787</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=19750401$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/1708285$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Perreault, Jean Pierre</creatorcontrib><creatorcontrib>Labuda, Damian</creatorcontrib><creatorcontrib>Usman, Nassim</creatorcontrib><creatorcontrib>Yang, Jing Hua</creatorcontrib><creatorcontrib>Cedergren, Robert</creatorcontrib><title>Relationship between 2'-hydroxyls and magnesium binding in the hammerhead RNA domain: a model for ribozyme catalysis</title><title>Biochemistry (Easton)</title><addtitle>Biochemistry</addtitle><description>The use of deoxyribonucleotide substitution in RNA (mixed RNA/DNA polymers) permits an evaluation of the role of 2'-hydroxyl groups in ribozyme catalysis. Specific deoxyribonucleotide substitution at G9 and A13 of the ribozyme decreases the catalytic activity (kcat) of the ribozyme by factors of 14 and 20, respectively. The reduction of the reaction rate concomitant with the absence of these 2'-OHs or the 2'-OH of the substrate U7 position can be partially compensated by increasing the Mg2+ concentration above 10 mM. The KMg of the all-RNA ribozyme is 5.3 mM, and the lack of either of the three influential 2'-OHs increases this value by a factor of approximately 3. These and other reaction constants for the ribozyme and the deoxy-substituted analogues have been determined by assuming a three-step mechanism. The data presented here provide the basis for the formulation of a molecular model of ribozyme activity.</description><subject>Base Sequence</subject><subject>Binding Sites</subject><subject>Biological and medical sciences</subject><subject>Catalysis</subject><subject>DNA - chemical synthesis</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Hydroxylation</subject><subject>Kinetics</subject><subject>Magnesium - chemistry</subject><subject>Mechanisms. Catalysis. Electron transfer. Models</subject><subject>Models, Genetic</subject><subject>Molecular biophysics</subject><subject>Molecular Sequence Data</subject><subject>Nucleic Acid Conformation</subject><subject>Physical chemistry in biology</subject><subject>RNA - chemical synthesis</subject><subject>RNA - chemistry</subject><subject>RNA, Catalytic - metabolism</subject><subject>Thermodynamics</subject><issn>0006-2960</issn><issn>1520-4995</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1991</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqF0TFv1TAQB3ALgcprYWJG8gIdUODsxLHTrapoQaoAlcJqXWKnzyW2H3YiGj49QXmiDEhMlvX_-eS7I-QZg9cMOHvTOgBeAgJvHpANExyKqmnEQ7IBgLrgTQ2PyWHOt8u1AlkdkAMmQXElNmS8sgOOLoa8dTva2vGHtYHy42I7mxTv5iFTDIZ6vAk2u8nT1gXjwg11gY5bS7fovU1bi4ZefTilJnp04YQi9dHYgfYx0eTa-HP2lnY44jBnl5-QRz0O2T7dn0fky_nb67N3xeXHi_dnp5cFVoqNhai5kWUPRhpjFW_6slGAsutM28haoICqUww5tFIxphTjXPBaKdX27fLSlkfk5Vp3l-L3yeZRe5c7OwwYbJyyViC4qIT6L2SiqWqm5AJfrbBLMedke71LzmOaNQP9exn6r2Us-vm-7NR6a-7tOv0lf7HPMXc49AlD5_I9a-TSIrDFFatzebR3f3JM33QtSyn09afP-kKwkn0tha4Xf7x67LK-jVMKy5T_-cNf6WqsHA</recordid><startdate>19910423</startdate><enddate>19910423</enddate><creator>Perreault, Jean Pierre</creator><creator>Labuda, Damian</creator><creator>Usman, Nassim</creator><creator>Yang, Jing Hua</creator><creator>Cedergren, Robert</creator><general>American Chemical Society</general><scope>BSCLL</scope><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7TM</scope><scope>7X8</scope></search><sort><creationdate>19910423</creationdate><title>Relationship between 2'-hydroxyls and magnesium binding in the hammerhead RNA domain: a model for ribozyme catalysis</title><author>Perreault, Jean Pierre ; Labuda, Damian ; Usman, Nassim ; Yang, Jing Hua ; Cedergren, Robert</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-a481t-562d73f0d7dde829f3980a7ccdb9765a504c81a20b781188122526888bfb2d7e3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1991</creationdate><topic>Base Sequence</topic><topic>Binding Sites</topic><topic>Biological and medical sciences</topic><topic>Catalysis</topic><topic>DNA - chemical synthesis</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Hydroxylation</topic><topic>Kinetics</topic><topic>Magnesium - chemistry</topic><topic>Mechanisms. Catalysis. Electron transfer. Models</topic><topic>Models, Genetic</topic><topic>Molecular biophysics</topic><topic>Molecular Sequence Data</topic><topic>Nucleic Acid Conformation</topic><topic>Physical chemistry in biology</topic><topic>RNA - chemical synthesis</topic><topic>RNA - chemistry</topic><topic>RNA, Catalytic - metabolism</topic><topic>Thermodynamics</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Perreault, Jean Pierre</creatorcontrib><creatorcontrib>Labuda, Damian</creatorcontrib><creatorcontrib>Usman, Nassim</creatorcontrib><creatorcontrib>Yang, Jing Hua</creatorcontrib><creatorcontrib>Cedergren, Robert</creatorcontrib><collection>Istex</collection><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Nucleic Acids Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Biochemistry (Easton)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Perreault, Jean Pierre</au><au>Labuda, Damian</au><au>Usman, Nassim</au><au>Yang, Jing Hua</au><au>Cedergren, Robert</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Relationship between 2'-hydroxyls and magnesium binding in the hammerhead RNA domain: a model for ribozyme catalysis</atitle><jtitle>Biochemistry (Easton)</jtitle><addtitle>Biochemistry</addtitle><date>1991-04-23</date><risdate>1991</risdate><volume>30</volume><issue>16</issue><spage>4020</spage><epage>4025</epage><pages>4020-4025</pages><issn>0006-2960</issn><eissn>1520-4995</eissn><abstract>The use of deoxyribonucleotide substitution in RNA (mixed RNA/DNA polymers) permits an evaluation of the role of 2'-hydroxyl groups in ribozyme catalysis. Specific deoxyribonucleotide substitution at G9 and A13 of the ribozyme decreases the catalytic activity (kcat) of the ribozyme by factors of 14 and 20, respectively. The reduction of the reaction rate concomitant with the absence of these 2'-OHs or the 2'-OH of the substrate U7 position can be partially compensated by increasing the Mg2+ concentration above 10 mM. The KMg of the all-RNA ribozyme is 5.3 mM, and the lack of either of the three influential 2'-OHs increases this value by a factor of approximately 3. These and other reaction constants for the ribozyme and the deoxy-substituted analogues have been determined by assuming a three-step mechanism. The data presented here provide the basis for the formulation of a molecular model of ribozyme activity.</abstract><cop>Washington, DC</cop><pub>American Chemical Society</pub><pmid>1708285</pmid><doi>10.1021/bi00230a029</doi><tpages>6</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0006-2960 |
| ispartof | Biochemistry (Easton), 1991-04, Vol.30 (16), p.4020-4025 |
| issn | 0006-2960 1520-4995 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_80525458 |
| source | MEDLINE; ACS Publications |
| subjects | Base Sequence Binding Sites Biological and medical sciences Catalysis DNA - chemical synthesis Fundamental and applied biological sciences. Psychology Hydroxylation Kinetics Magnesium - chemistry Mechanisms. Catalysis. Electron transfer. Models Models, Genetic Molecular biophysics Molecular Sequence Data Nucleic Acid Conformation Physical chemistry in biology RNA - chemical synthesis RNA - chemistry RNA, Catalytic - metabolism Thermodynamics |
| title | Relationship between 2'-hydroxyls and magnesium binding in the hammerhead RNA domain: a model for ribozyme catalysis |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-08T13%3A34%3A17IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Relationship%20between%202'-hydroxyls%20and%20magnesium%20binding%20in%20the%20hammerhead%20RNA%20domain:%20a%20model%20for%20ribozyme%20catalysis&rft.jtitle=Biochemistry%20(Easton)&rft.au=Perreault,%20Jean%20Pierre&rft.date=1991-04-23&rft.volume=30&rft.issue=16&rft.spage=4020&rft.epage=4025&rft.pages=4020-4025&rft.issn=0006-2960&rft.eissn=1520-4995&rft_id=info:doi/10.1021/bi00230a029&rft_dat=%3Cproquest_cross%3E80525458%3C/proquest_cross%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=15946187&rft_id=info:pmid/1708285&rfr_iscdi=true |