Ribosome release factor RF4 and termination factor RF3 are involved in dissociation of peptidyl-tRNA from the ribosome
Peptidyl‐tRNA dissociation from ribosomes is an energetically costly but apparently inevitable process that accompanies normal protein synthesis. The drop‐off products of these events are hydrolysed by peptidyl‐tRNA hydrolase. Mutant selections have been made to identify genes involved in the drop‐o...
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| Veröffentlicht in: | The EMBO journal 1998-02, Vol.17 (3), p.808-816 |
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| creator | Heurgué-Hamard, Valérie Karimi, Reza Mora, Liliana MacDougall, Jane Leboeuf, Céline Grentzmann, Guido Ehrenberg, Måns Buckingham, Richard H. |
| description | Peptidyl‐tRNA dissociation from ribosomes is an energetically costly but apparently inevitable process that accompanies normal protein synthesis. The drop‐off products of these events are hydrolysed by peptidyl‐tRNA hydrolase. Mutant selections have been made to identify genes involved in the drop‐off of peptidyl‐tRNA, using a thermosensitive peptidyl‐tRNA hydrolase mutant in Escherichia coli. Transposon insertions upstream of the frr gene, which encodes RF4 (ribosome release or recycling factor), restored growth to this mutant. The insertions impaired expression of the frr gene. Mutations inactivating prfC, encoding RF3 (release factor 3), displayed a similar phenotype. Conversely, production of RF4 from a plasmid increased the thermosensitivity of the peptidyl‐tRNA hydrolase mutant. In vitro measurements of peptidyl‐tRNA release from ribosomes paused at stop signals or sense codons confirmed that RF3 and RF4 were able to stimulate peptidyl‐tRNA release from ribosomes, and showed that this action of RF4 required the presence of translocation factor EF2, known to be needed for the function of RF4 in ribosome recycling. When present together, the three factors were able to stimulate release up to 12‐fold. It is suggested that RF4 may displace peptidyl‐tRNA from the ribosome in a manner related to its proposed function in removing deacylated tRNA during ribosome recycling. |
| doi_str_mv | 10.1093/emboj/17.3.808 |
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The drop‐off products of these events are hydrolysed by peptidyl‐tRNA hydrolase. Mutant selections have been made to identify genes involved in the drop‐off of peptidyl‐tRNA, using a thermosensitive peptidyl‐tRNA hydrolase mutant in Escherichia coli. Transposon insertions upstream of the frr gene, which encodes RF4 (ribosome release or recycling factor), restored growth to this mutant. The insertions impaired expression of the frr gene. Mutations inactivating prfC, encoding RF3 (release factor 3), displayed a similar phenotype. Conversely, production of RF4 from a plasmid increased the thermosensitivity of the peptidyl‐tRNA hydrolase mutant. In vitro measurements of peptidyl‐tRNA release from ribosomes paused at stop signals or sense codons confirmed that RF3 and RF4 were able to stimulate peptidyl‐tRNA release from ribosomes, and showed that this action of RF4 required the presence of translocation factor EF2, known to be needed for the function of RF4 in ribosome recycling. When present together, the three factors were able to stimulate release up to 12‐fold. It is suggested that RF4 may displace peptidyl‐tRNA from the ribosome in a manner related to its proposed function in removing deacylated tRNA during ribosome recycling.</description><identifier>ISSN: 0261-4189</identifier><identifier>ISSN: 1460-2075</identifier><identifier>EISSN: 1460-2075</identifier><identifier>DOI: 10.1093/emboj/17.3.808</identifier><identifier>PMID: 9451005</identifier><language>eng</language><publisher>Chichester, UK: John Wiley & Sons, Ltd</publisher><subject>Bacterial Proteins - genetics ; Bacterial Proteins - metabolism ; Carboxylic Ester Hydrolases - genetics ; Culture Media ; DNA Transposable Elements - genetics ; Escherichia coli - genetics ; Escherichia coli - growth & development ; Gene Expression Regulation, Bacterial ; Genes, Bacterial - genetics ; Guanosine Triphosphate - physiology ; Mutation - genetics ; Mutation - physiology ; Peptide Chain Termination, Translational - physiology ; Peptide Elongation Factor 2 ; Peptide Elongation Factors - metabolism ; Peptide Termination Factors - genetics ; Peptide Termination Factors - metabolism ; protein synthesis ; Proteins ; release factors ; RF3 ; RF4 ; Ribosomal Proteins ; ribosome ; Ribosomes - metabolism ; RNA, Transfer, Amino Acyl - metabolism ; Suppression, Genetic ; Temperature</subject><ispartof>The EMBO journal, 1998-02, Vol.17 (3), p.808-816</ispartof><rights>Copyright © 1998 European Molecular Biology Organization</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c5686-456378b18b4cd957fd422895a7af81ac34a1eb842ab8802e46b069e8760da1733</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC1170429/pdf/$$EPDF$$P50$$Gpubmedcentral$$H</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC1170429/$$EHTML$$P50$$Gpubmedcentral$$H</linktohtml><link.rule.ids>230,314,723,776,780,881,1411,1427,27901,27902,45550,45551,46384,46808,53766,53768</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/9451005$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Heurgué-Hamard, Valérie</creatorcontrib><creatorcontrib>Karimi, Reza</creatorcontrib><creatorcontrib>Mora, Liliana</creatorcontrib><creatorcontrib>MacDougall, Jane</creatorcontrib><creatorcontrib>Leboeuf, Céline</creatorcontrib><creatorcontrib>Grentzmann, Guido</creatorcontrib><creatorcontrib>Ehrenberg, Måns</creatorcontrib><creatorcontrib>Buckingham, Richard H.</creatorcontrib><title>Ribosome release factor RF4 and termination factor RF3 are involved in dissociation of peptidyl-tRNA from the ribosome</title><title>The EMBO journal</title><addtitle>EMBO J</addtitle><description>Peptidyl‐tRNA dissociation from ribosomes is an energetically costly but apparently inevitable process that accompanies normal protein synthesis. The drop‐off products of these events are hydrolysed by peptidyl‐tRNA hydrolase. Mutant selections have been made to identify genes involved in the drop‐off of peptidyl‐tRNA, using a thermosensitive peptidyl‐tRNA hydrolase mutant in Escherichia coli. Transposon insertions upstream of the frr gene, which encodes RF4 (ribosome release or recycling factor), restored growth to this mutant. The insertions impaired expression of the frr gene. Mutations inactivating prfC, encoding RF3 (release factor 3), displayed a similar phenotype. Conversely, production of RF4 from a plasmid increased the thermosensitivity of the peptidyl‐tRNA hydrolase mutant. In vitro measurements of peptidyl‐tRNA release from ribosomes paused at stop signals or sense codons confirmed that RF3 and RF4 were able to stimulate peptidyl‐tRNA release from ribosomes, and showed that this action of RF4 required the presence of translocation factor EF2, known to be needed for the function of RF4 in ribosome recycling. When present together, the three factors were able to stimulate release up to 12‐fold. It is suggested that RF4 may displace peptidyl‐tRNA from the ribosome in a manner related to its proposed function in removing deacylated tRNA during ribosome recycling.</description><subject>Bacterial Proteins - genetics</subject><subject>Bacterial Proteins - metabolism</subject><subject>Carboxylic Ester Hydrolases - genetics</subject><subject>Culture Media</subject><subject>DNA Transposable Elements - genetics</subject><subject>Escherichia coli - genetics</subject><subject>Escherichia coli - growth & development</subject><subject>Gene Expression Regulation, Bacterial</subject><subject>Genes, Bacterial - genetics</subject><subject>Guanosine Triphosphate - physiology</subject><subject>Mutation - genetics</subject><subject>Mutation - physiology</subject><subject>Peptide Chain Termination, Translational - physiology</subject><subject>Peptide Elongation Factor 2</subject><subject>Peptide Elongation Factors - metabolism</subject><subject>Peptide Termination Factors - genetics</subject><subject>Peptide Termination Factors - metabolism</subject><subject>protein synthesis</subject><subject>Proteins</subject><subject>release factors</subject><subject>RF3</subject><subject>RF4</subject><subject>Ribosomal Proteins</subject><subject>ribosome</subject><subject>Ribosomes - metabolism</subject><subject>RNA, Transfer, Amino Acyl - metabolism</subject><subject>Suppression, Genetic</subject><subject>Temperature</subject><issn>0261-4189</issn><issn>1460-2075</issn><issn>1460-2075</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1998</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkc9v0zAcxS0EGmVw5YbkE7d0dvz7grRN6woaQ-pgHC0n-YZ5JHGx3UL_ezJSFTjtZFvvvc_3az2EXlMyp8SwE-ircH9C1ZzNNdFP0IxySYqSKPEUzUgpacGpNs_Ri5TuCSFCK3qEjgwXdHzM0Hblq5BCDzhCBy4Bbl2dQ8SrBcduaHCG2PvBZR-GvxLDLgL2wzZ0W2jGC258SqH2ky-0eA3r7JtdV-TV9SluY-hxvhuH7Ke9RM9a1yV4tT-P0ZfFxefzZXH16fL9-elVUQupZcGFZEpXVFe8boxQbcPLUhvhlGs1dTXjjkKleekqrUkJXFZEGtBKksZRxdgxejdx15uqh6aGIUfX2XX0vYs7G5y3_yuDv7PfwtZSqggvzQh4uwfE8GMDKdvepxq6zg0QNskqI5XglD1qpLLUhLIH4nwy1jGkFKE9bEOJfajU_qnUUmWZHSsdA2_-_cPBvu9w1NWk__Qd7B6h2YuPZx-UMGRcfEwWU9KnDL8OSRe_W6mYEvbr9aW9uV0sz5a3N9aw3x9Gv4U</recordid><startdate>19980202</startdate><enddate>19980202</enddate><creator>Heurgué-Hamard, Valérie</creator><creator>Karimi, Reza</creator><creator>Mora, Liliana</creator><creator>MacDougall, Jane</creator><creator>Leboeuf, Céline</creator><creator>Grentzmann, Guido</creator><creator>Ehrenberg, Måns</creator><creator>Buckingham, Richard H.</creator><general>John Wiley & Sons, Ltd</general><scope>BSCLL</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><scope>5PM</scope></search><sort><creationdate>19980202</creationdate><title>Ribosome release factor RF4 and termination factor RF3 are involved in dissociation of peptidyl-tRNA from the ribosome</title><author>Heurgué-Hamard, Valérie ; Karimi, Reza ; Mora, Liliana ; MacDougall, Jane ; Leboeuf, Céline ; Grentzmann, Guido ; Ehrenberg, Måns ; Buckingham, Richard H.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c5686-456378b18b4cd957fd422895a7af81ac34a1eb842ab8802e46b069e8760da1733</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1998</creationdate><topic>Bacterial Proteins - genetics</topic><topic>Bacterial Proteins - metabolism</topic><topic>Carboxylic Ester Hydrolases - genetics</topic><topic>Culture Media</topic><topic>DNA Transposable Elements - genetics</topic><topic>Escherichia coli - genetics</topic><topic>Escherichia coli - growth & development</topic><topic>Gene Expression Regulation, Bacterial</topic><topic>Genes, Bacterial - genetics</topic><topic>Guanosine Triphosphate - physiology</topic><topic>Mutation - genetics</topic><topic>Mutation - physiology</topic><topic>Peptide Chain Termination, Translational - physiology</topic><topic>Peptide Elongation Factor 2</topic><topic>Peptide Elongation Factors - metabolism</topic><topic>Peptide Termination Factors - genetics</topic><topic>Peptide Termination Factors - metabolism</topic><topic>protein synthesis</topic><topic>Proteins</topic><topic>release factors</topic><topic>RF3</topic><topic>RF4</topic><topic>Ribosomal Proteins</topic><topic>ribosome</topic><topic>Ribosomes - metabolism</topic><topic>RNA, Transfer, Amino Acyl - metabolism</topic><topic>Suppression, Genetic</topic><topic>Temperature</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Heurgué-Hamard, Valérie</creatorcontrib><creatorcontrib>Karimi, Reza</creatorcontrib><creatorcontrib>Mora, Liliana</creatorcontrib><creatorcontrib>MacDougall, Jane</creatorcontrib><creatorcontrib>Leboeuf, Céline</creatorcontrib><creatorcontrib>Grentzmann, Guido</creatorcontrib><creatorcontrib>Ehrenberg, Måns</creatorcontrib><creatorcontrib>Buckingham, Richard H.</creatorcontrib><collection>Istex</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><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>Heurgué-Hamard, Valérie</au><au>Karimi, Reza</au><au>Mora, Liliana</au><au>MacDougall, Jane</au><au>Leboeuf, Céline</au><au>Grentzmann, Guido</au><au>Ehrenberg, Måns</au><au>Buckingham, Richard H.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Ribosome release factor RF4 and termination factor RF3 are involved in dissociation of peptidyl-tRNA from the ribosome</atitle><jtitle>The EMBO journal</jtitle><addtitle>EMBO J</addtitle><date>1998-02-02</date><risdate>1998</risdate><volume>17</volume><issue>3</issue><spage>808</spage><epage>816</epage><pages>808-816</pages><issn>0261-4189</issn><issn>1460-2075</issn><eissn>1460-2075</eissn><abstract>Peptidyl‐tRNA dissociation from ribosomes is an energetically costly but apparently inevitable process that accompanies normal protein synthesis. The drop‐off products of these events are hydrolysed by peptidyl‐tRNA hydrolase. Mutant selections have been made to identify genes involved in the drop‐off of peptidyl‐tRNA, using a thermosensitive peptidyl‐tRNA hydrolase mutant in Escherichia coli. Transposon insertions upstream of the frr gene, which encodes RF4 (ribosome release or recycling factor), restored growth to this mutant. The insertions impaired expression of the frr gene. Mutations inactivating prfC, encoding RF3 (release factor 3), displayed a similar phenotype. Conversely, production of RF4 from a plasmid increased the thermosensitivity of the peptidyl‐tRNA hydrolase mutant. In vitro measurements of peptidyl‐tRNA release from ribosomes paused at stop signals or sense codons confirmed that RF3 and RF4 were able to stimulate peptidyl‐tRNA release from ribosomes, and showed that this action of RF4 required the presence of translocation factor EF2, known to be needed for the function of RF4 in ribosome recycling. When present together, the three factors were able to stimulate release up to 12‐fold. It is suggested that RF4 may displace peptidyl‐tRNA from the ribosome in a manner related to its proposed function in removing deacylated tRNA during ribosome recycling.</abstract><cop>Chichester, UK</cop><pub>John Wiley & Sons, Ltd</pub><pmid>9451005</pmid><doi>10.1093/emboj/17.3.808</doi><tpages>9</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Bacterial Proteins - genetics Bacterial Proteins - metabolism Carboxylic Ester Hydrolases - genetics Culture Media DNA Transposable Elements - genetics Escherichia coli - genetics Escherichia coli - growth & development Gene Expression Regulation, Bacterial Genes, Bacterial - genetics Guanosine Triphosphate - physiology Mutation - genetics Mutation - physiology Peptide Chain Termination, Translational - physiology Peptide Elongation Factor 2 Peptide Elongation Factors - metabolism Peptide Termination Factors - genetics Peptide Termination Factors - metabolism protein synthesis Proteins release factors RF3 RF4 Ribosomal Proteins ribosome Ribosomes - metabolism RNA, Transfer, Amino Acyl - metabolism Suppression, Genetic Temperature |
| title | Ribosome release factor RF4 and termination factor RF3 are involved in dissociation of peptidyl-tRNA from the ribosome |
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