Antisense RNA‐mediated transcriptional attenuation occurs faster than stable antisense/target RNA pairing: an in vitro study of plasmid pIP501
Antisense RNA‐mediated transcriptional attenuation is the mode of replication control of several plasmids, among them pIP501. This mechanism implies that the repR mRNAs can fold into two mutually exclusive structures. The formation of one of these structures is induced by binding of the antisense RN...
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| Veröffentlicht in: | The EMBO journal 1994-08, Vol.13 (15), p.3599-3607 |
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| description | Antisense RNA‐mediated transcriptional attenuation is the mode of replication control of several plasmids, among them pIP501. This mechanism implies that the repR mRNAs can fold into two mutually exclusive structures. The formation of one of these structures is induced by binding of the antisense RNA and results in premature termination. Since the fate of the nascent mRNA transcripts depends on the binding rate of the antisense RNA to its target, the control is kinetic. We have studied the antisense RNA, RNAIII, and target RNA, RNAII, whose interaction determines the replication frequency of plasmid pIP501. RNA secondary structures were analyzed using structure‐specific RNases. RNA binding was studied in vitro with normal size and truncated RNAIII species. An in vitro single‐round attenuation assay was developed that permits qualitative and quantitative assessment of inhibition by RNAIII. The effect of varying concentrations of RNAIII species on attenuation was tested and inhibition rate constants were calculated. The inhibition rate constants were at least 10 times higher than the pairing rate constants. Thus, steps preceding stable RNA duplex formation are sufficient to induce RNAIII‐dependent termination of nascent RNAII transcripts. |
| doi_str_mv | 10.1002/j.1460-2075.1994.tb06667.x |
| format | Article |
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This mechanism implies that the repR mRNAs can fold into two mutually exclusive structures. The formation of one of these structures is induced by binding of the antisense RNA and results in premature termination. Since the fate of the nascent mRNA transcripts depends on the binding rate of the antisense RNA to its target, the control is kinetic. We have studied the antisense RNA, RNAIII, and target RNA, RNAII, whose interaction determines the replication frequency of plasmid pIP501. RNA secondary structures were analyzed using structure‐specific RNases. RNA binding was studied in vitro with normal size and truncated RNAIII species. An in vitro single‐round attenuation assay was developed that permits qualitative and quantitative assessment of inhibition by RNAIII. The effect of varying concentrations of RNAIII species on attenuation was tested and inhibition rate constants were calculated. The inhibition rate constants were at least 10 times higher than the pairing rate constants. Thus, steps preceding stable RNA duplex formation are sufficient to induce RNAIII‐dependent termination of nascent RNAII transcripts.</description><identifier>ISSN: 0261-4189</identifier><identifier>EISSN: 1460-2075</identifier><identifier>DOI: 10.1002/j.1460-2075.1994.tb06667.x</identifier><identifier>PMID: 7520390</identifier><identifier>CODEN: EMJODG</identifier><language>eng</language><publisher>London: Nature Publishing Group</publisher><subject>Bacteriology ; Base Sequence ; Biological and medical sciences ; DNA Replication - physiology ; Fundamental and applied biological sciences. 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This mechanism implies that the repR mRNAs can fold into two mutually exclusive structures. The formation of one of these structures is induced by binding of the antisense RNA and results in premature termination. Since the fate of the nascent mRNA transcripts depends on the binding rate of the antisense RNA to its target, the control is kinetic. We have studied the antisense RNA, RNAIII, and target RNA, RNAII, whose interaction determines the replication frequency of plasmid pIP501. RNA secondary structures were analyzed using structure‐specific RNases. RNA binding was studied in vitro with normal size and truncated RNAIII species. An in vitro single‐round attenuation assay was developed that permits qualitative and quantitative assessment of inhibition by RNAIII. The effect of varying concentrations of RNAIII species on attenuation was tested and inhibition rate constants were calculated. The inhibition rate constants were at least 10 times higher than the pairing rate constants. Thus, steps preceding stable RNA duplex formation are sufficient to induce RNAIII‐dependent termination of nascent RNAII transcripts.</description><subject>Bacteriology</subject><subject>Base Sequence</subject><subject>Biological and medical sciences</subject><subject>DNA Replication - physiology</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Genetics</subject><subject>Kinetics</subject><subject>Microbiology</subject><subject>Models, Molecular</subject><subject>Molecular Sequence Data</subject><subject>Nucleic Acid Conformation</subject><subject>Plasmids - genetics</subject><subject>RNA - chemistry</subject><subject>RNA - metabolism</subject><subject>RNA, Antisense - chemistry</subject><subject>RNA, Antisense - metabolism</subject><subject>RNA, Antisense - physiology</subject><subject>Transcription, Genetic - physiology</subject><issn>0261-4189</issn><issn>1460-2075</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1994</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqVkc9u1DAQxi0EKkvhEZAshLhtasdJnFTisFQFisofIThbE2ey9SrrpLZTujceoc_Ik-CwYQUnxMm2vu8bz8yPkGecJZyx9GST8Kxgy5TJPOFVlSWhZkVRyOT2HlkcpPtkwdKCLzNeVg_JI-83jLG8lPyIHMk8ZaJiC3K3ssF4tB7p5w-rH9_vttgYCNjQ4MB67cwQTG-hoxAC2hGmF-21Hp2nLfiAjoYrsNQHqDuk8LvcSQC3xjBVpQMYZ-z6NKrUWHpjgutjYGx2tG_p0IHfmoYOF59yxh-TBy10Hp_M5zH5-vr8y9nb5eXHNxdnq8ulzkUplyITGa9KobGsRF6nmpeMtSKtodVxFQUIXmLFsGxQ1pnM6hREqpnGuCguGYpj8nJfdxjrOLNGGwfu1ODMFtxO9WDU34o1V2rd3yhR5WmRx_yLOe_66xF9UFvjNXYdWOxHr2TkEf-S_zTyoiyLqsii8XRv1K733mF7aIYzNXFXGzXBVRNcNXFXM3d1G8NP_xznEJ1BR_35rIPX0LURrjb-YMtSIXI52VZ72zfT4e4_GlDn71-9-3UXPwF4Kc66</recordid><startdate>199408</startdate><enddate>199408</enddate><creator>Brantl, S.</creator><creator>Wagner, E.G.</creator><general>Nature Publishing Group</general><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>7QL</scope><scope>7TM</scope><scope>C1K</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>199408</creationdate><title>Antisense RNA‐mediated transcriptional attenuation occurs faster than stable antisense/target RNA pairing: an in vitro study of plasmid pIP501</title><author>Brantl, S. ; Wagner, E.G.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c5387-34341983ce8935b2c1800f32bafc6666a318e90e8de7b474b2a32c0ce066170e3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1994</creationdate><topic>Bacteriology</topic><topic>Base Sequence</topic><topic>Biological and medical sciences</topic><topic>DNA Replication - physiology</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Genetics</topic><topic>Kinetics</topic><topic>Microbiology</topic><topic>Models, Molecular</topic><topic>Molecular Sequence Data</topic><topic>Nucleic Acid Conformation</topic><topic>Plasmids - genetics</topic><topic>RNA - chemistry</topic><topic>RNA - metabolism</topic><topic>RNA, Antisense - chemistry</topic><topic>RNA, Antisense - metabolism</topic><topic>RNA, Antisense - physiology</topic><topic>Transcription, Genetic - physiology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Brantl, S.</creatorcontrib><creatorcontrib>Wagner, E.G.</creatorcontrib><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>Bacteriology Abstracts (Microbiology B)</collection><collection>Nucleic Acids Abstracts</collection><collection>Environmental Sciences and Pollution Management</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>The EMBO journal</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Brantl, S.</au><au>Wagner, E.G.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Antisense RNA‐mediated transcriptional attenuation occurs faster than stable antisense/target RNA pairing: an in vitro study of plasmid pIP501</atitle><jtitle>The EMBO journal</jtitle><addtitle>EMBO J</addtitle><date>1994-08</date><risdate>1994</risdate><volume>13</volume><issue>15</issue><spage>3599</spage><epage>3607</epage><pages>3599-3607</pages><issn>0261-4189</issn><eissn>1460-2075</eissn><coden>EMJODG</coden><abstract>Antisense RNA‐mediated transcriptional attenuation is the mode of replication control of several plasmids, among them pIP501. This mechanism implies that the repR mRNAs can fold into two mutually exclusive structures. The formation of one of these structures is induced by binding of the antisense RNA and results in premature termination. Since the fate of the nascent mRNA transcripts depends on the binding rate of the antisense RNA to its target, the control is kinetic. We have studied the antisense RNA, RNAIII, and target RNA, RNAII, whose interaction determines the replication frequency of plasmid pIP501. RNA secondary structures were analyzed using structure‐specific RNases. RNA binding was studied in vitro with normal size and truncated RNAIII species. An in vitro single‐round attenuation assay was developed that permits qualitative and quantitative assessment of inhibition by RNAIII. The effect of varying concentrations of RNAIII species on attenuation was tested and inhibition rate constants were calculated. The inhibition rate constants were at least 10 times higher than the pairing rate constants. Thus, steps preceding stable RNA duplex formation are sufficient to induce RNAIII‐dependent termination of nascent RNAII transcripts.</abstract><cop>London</cop><pub>Nature Publishing Group</pub><pmid>7520390</pmid><doi>10.1002/j.1460-2075.1994.tb06667.x</doi><tpages>9</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Bacteriology Base Sequence Biological and medical sciences DNA Replication - physiology Fundamental and applied biological sciences. Psychology Genetics Kinetics Microbiology Models, Molecular Molecular Sequence Data Nucleic Acid Conformation Plasmids - genetics RNA - chemistry RNA - metabolism RNA, Antisense - chemistry RNA, Antisense - metabolism RNA, Antisense - physiology Transcription, Genetic - physiology |
| title | Antisense RNA‐mediated transcriptional attenuation occurs faster than stable antisense/target RNA pairing: an in vitro study of plasmid pIP501 |
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