Quantitative Determination that One of Two Potential RNA-Binding Domains of the A Protein Component of the U1 Small Nuclear Ribonucleoprotein Complex Binds with High Affinity to Stem-Loop II of U1 RNA
Many RNA-associated proteins contain a ribonucleoprotein (RNP) consensus octamer encompassed by a conserved 80 amino acid sequence, which we have termed an RNA recognition motif (RRM). RRM family members contain either one (class I) or multiple (class II) copies of this motif. We report here that a...
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| Veröffentlicht in: | Proceedings of the National Academy of Sciences - PNAS 1990-08, Vol.87 (16), p.6393-6397 |
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| creator | Lutz-Freyermuth, Carol Query, Charles C. Keene, Jack D. |
| description | Many RNA-associated proteins contain a ribonucleoprotein (RNP) consensus octamer encompassed by a conserved 80 amino acid sequence, which we have termed an RNA recognition motif (RRM). RRM family members contain either one (class I) or multiple (class II) copies of this motif. We report here that a class II component of the U1 small nuclear RNP (snRNP), the A protein of U1 snRNP (U1snRNP-A), contains two RRMs (RRM1 and -2), yet has only one binding domain (RRM1) that interacts specifically with stem-loop II of U1 RNA. Quantitative analysis of binding affinities of fragments of U1snRNP-A demonstrated that an 86-amino acid polypeptide was competent to bind to U1 RNA with an affinity comparable to that of the full-length protein (Kd≈ 80 nM). The carboxyl-terminal RRM2 of U1snRNP-A did not bind to U1 RNA and may recognize an unidentified heterologous RNA. We propose that class II proteins may function as bridges between RNA components of RNP complexes such as the spliceosome. |
| doi_str_mv | 10.1073/pnas.87.16.6393 |
| format | Article |
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RRM family members contain either one (class I) or multiple (class II) copies of this motif. We report here that a class II component of the U1 small nuclear RNP (snRNP), the A protein of U1 snRNP (U1snRNP-A), contains two RRMs (RRM1 and -2), yet has only one binding domain (RRM1) that interacts specifically with stem-loop II of U1 RNA. Quantitative analysis of binding affinities of fragments of U1snRNP-A demonstrated that an 86-amino acid polypeptide was competent to bind to U1 RNA with an affinity comparable to that of the full-length protein (Kd≈ 80 nM). The carboxyl-terminal RRM2 of U1snRNP-A did not bind to U1 RNA and may recognize an unidentified heterologous RNA. We propose that class II proteins may function as bridges between RNA components of RNP complexes such as the spliceosome.</description><identifier>ISSN: 0027-8424</identifier><identifier>EISSN: 1091-6490</identifier><identifier>DOI: 10.1073/pnas.87.16.6393</identifier><identifier>PMID: 1696729</identifier><identifier>CODEN: PNASA6</identifier><language>eng</language><publisher>Washington, DC: National Academy of Sciences of the United States of America</publisher><subject>Amino acids ; Analytical, structural and metabolic biochemistry ; Antibodies ; Binding Sites ; Biochemistry ; Biological and medical sciences ; Electrophoresis, Polyacrylamide Gel ; Epitopes - analysis ; Fundamental and applied biological sciences. Psychology ; Gels ; Gene Library ; HeLa cells ; HeLa Cells - metabolism ; Humans ; Kinetics ; Macromolecular Substances ; Mutation ; Nucleic Acid Conformation ; Nucleic acids ; Oligonucleotide Probes ; Protein Biosynthesis ; Proteins ; Ribonucleoproteins - genetics ; Ribonucleoproteins - isolation & purification ; Ribonucleoproteins - metabolism ; Ribonucleoproteins, Small Nuclear ; RNA ; Rna, ribonucleoproteins ; RNA, Small Nuclear - metabolism ; Small nuclear ribonucleoproteins ; Splicing ; Transcription, Genetic</subject><ispartof>Proceedings of the National Academy of Sciences - PNAS, 1990-08, Vol.87 (16), p.6393-6397</ispartof><rights>1991 INIST-CNRS</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c4383-5e45f6eb24c4f526af481bbcef7329f20bbb61fb2f242b876b5e836271cb8dfc3</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Uhttp://www.pnas.org/content/87/16.cover.gif</thumbnail><linktopdf>$$Uhttps://www.jstor.org/stable/pdf/2356214$$EPDF$$P50$$Gjstor$$H</linktopdf><linktohtml>$$Uhttps://www.jstor.org/stable/2356214$$EHTML$$P50$$Gjstor$$H</linktohtml><link.rule.ids>230,314,723,776,780,799,881,27901,27902,53766,53768,57992,58225</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=19436381$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/1696729$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Lutz-Freyermuth, Carol</creatorcontrib><creatorcontrib>Query, Charles C.</creatorcontrib><creatorcontrib>Keene, Jack D.</creatorcontrib><title>Quantitative Determination that One of Two Potential RNA-Binding Domains of the A Protein Component of the U1 Small Nuclear Ribonucleoprotein Complex Binds with High Affinity to Stem-Loop II of U1 RNA</title><title>Proceedings of the National Academy of Sciences - PNAS</title><addtitle>Proc Natl Acad Sci U S A</addtitle><description>Many RNA-associated proteins contain a ribonucleoprotein (RNP) consensus octamer encompassed by a conserved 80 amino acid sequence, which we have termed an RNA recognition motif (RRM). RRM family members contain either one (class I) or multiple (class II) copies of this motif. We report here that a class II component of the U1 small nuclear RNP (snRNP), the A protein of U1 snRNP (U1snRNP-A), contains two RRMs (RRM1 and -2), yet has only one binding domain (RRM1) that interacts specifically with stem-loop II of U1 RNA. Quantitative analysis of binding affinities of fragments of U1snRNP-A demonstrated that an 86-amino acid polypeptide was competent to bind to U1 RNA with an affinity comparable to that of the full-length protein (Kd≈ 80 nM). The carboxyl-terminal RRM2 of U1snRNP-A did not bind to U1 RNA and may recognize an unidentified heterologous RNA. We propose that class II proteins may function as bridges between RNA components of RNP complexes such as the spliceosome.</description><subject>Amino acids</subject><subject>Analytical, structural and metabolic biochemistry</subject><subject>Antibodies</subject><subject>Binding Sites</subject><subject>Biochemistry</subject><subject>Biological and medical sciences</subject><subject>Electrophoresis, Polyacrylamide Gel</subject><subject>Epitopes - analysis</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gels</subject><subject>Gene Library</subject><subject>HeLa cells</subject><subject>HeLa Cells - metabolism</subject><subject>Humans</subject><subject>Kinetics</subject><subject>Macromolecular Substances</subject><subject>Mutation</subject><subject>Nucleic Acid Conformation</subject><subject>Nucleic acids</subject><subject>Oligonucleotide Probes</subject><subject>Protein Biosynthesis</subject><subject>Proteins</subject><subject>Ribonucleoproteins - genetics</subject><subject>Ribonucleoproteins - isolation & purification</subject><subject>Ribonucleoproteins - metabolism</subject><subject>Ribonucleoproteins, Small Nuclear</subject><subject>RNA</subject><subject>Rna, ribonucleoproteins</subject><subject>RNA, Small Nuclear - metabolism</subject><subject>Small nuclear ribonucleoproteins</subject><subject>Splicing</subject><subject>Transcription, Genetic</subject><issn>0027-8424</issn><issn>1091-6490</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1990</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkktv1DAUhSMEKkNhzQaQN8AqU7_ixFI3wxRopVFb-lhbTsaeceXYIXb6-If8LBzN9LWBlW2d75x7LZ0se4_gFMGS7HVOhmlVThGbMsLJi2yCIEc5oxy-zCYQ4jKvKKavszchXEEIeVHBnWwHMc5KzCfZn1-DdNFEGc21Agcqqr41Lr28A3EtIzhxCngNLm48OPVRJVZacHY8y78ZtzRuBQ58K40LIxTXCszAaZ8448Dct513yXEvXSJw3kprwfHQWCV7cGZq78a77554rLoFY3gANyauwaFZrcFMa-NMvAPRg_Oo2nzhfQeOjsboFJv2eZu90tIG9W577maXP75fzA_zxcnPo_lskTeUVCQvFC00UzWmDdUFZlLTCtV1o3RJMNcY1nXNkK6xxhTXVcnqQlWE4RI1dbXUDdnN9je53VC3atmk__XSiq43rezvhJdGPFecWYuVvxYFLShM9i9be-9_DypE0ZrQKGulU34IouScIlKx_4Ko4BwWiCRwbwM2vQ-hV_phFwTF2BExdkRUpUBMjB1Jjo9Pv_DIb0qR9M9bXYZGWt1L15jwiHFKGKlQ4j5tuXHAvfxs0Nd_AkIP1kZ1GxP5YUNehej7BxSTgmFEyV9pNum5</recordid><startdate>19900801</startdate><enddate>19900801</enddate><creator>Lutz-Freyermuth, Carol</creator><creator>Query, Charles C.</creator><creator>Keene, Jack D.</creator><general>National Academy of Sciences of the United States of America</general><general>National Acad Sciences</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>7TM</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>19900801</creationdate><title>Quantitative Determination that One of Two Potential RNA-Binding Domains of the A Protein Component of the U1 Small Nuclear Ribonucleoprotein Complex Binds with High Affinity to Stem-Loop II of U1 RNA</title><author>Lutz-Freyermuth, Carol ; Query, Charles C. ; Keene, Jack D.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4383-5e45f6eb24c4f526af481bbcef7329f20bbb61fb2f242b876b5e836271cb8dfc3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1990</creationdate><topic>Amino acids</topic><topic>Analytical, structural and metabolic biochemistry</topic><topic>Antibodies</topic><topic>Binding Sites</topic><topic>Biochemistry</topic><topic>Biological and medical sciences</topic><topic>Electrophoresis, Polyacrylamide Gel</topic><topic>Epitopes - analysis</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Gels</topic><topic>Gene Library</topic><topic>HeLa cells</topic><topic>HeLa Cells - metabolism</topic><topic>Humans</topic><topic>Kinetics</topic><topic>Macromolecular Substances</topic><topic>Mutation</topic><topic>Nucleic Acid Conformation</topic><topic>Nucleic acids</topic><topic>Oligonucleotide Probes</topic><topic>Protein Biosynthesis</topic><topic>Proteins</topic><topic>Ribonucleoproteins - genetics</topic><topic>Ribonucleoproteins - isolation & purification</topic><topic>Ribonucleoproteins - metabolism</topic><topic>Ribonucleoproteins, Small Nuclear</topic><topic>RNA</topic><topic>Rna, ribonucleoproteins</topic><topic>RNA, Small Nuclear - metabolism</topic><topic>Small nuclear ribonucleoproteins</topic><topic>Splicing</topic><topic>Transcription, Genetic</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Lutz-Freyermuth, Carol</creatorcontrib><creatorcontrib>Query, Charles C.</creatorcontrib><creatorcontrib>Keene, Jack D.</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>Nucleic Acids Abstracts</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Proceedings of the National Academy of Sciences - PNAS</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Lutz-Freyermuth, Carol</au><au>Query, Charles C.</au><au>Keene, Jack D.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Quantitative Determination that One of Two Potential RNA-Binding Domains of the A Protein Component of the U1 Small Nuclear Ribonucleoprotein Complex Binds with High Affinity to Stem-Loop II of U1 RNA</atitle><jtitle>Proceedings of the National Academy of Sciences - PNAS</jtitle><addtitle>Proc Natl Acad Sci U S A</addtitle><date>1990-08-01</date><risdate>1990</risdate><volume>87</volume><issue>16</issue><spage>6393</spage><epage>6397</epage><pages>6393-6397</pages><issn>0027-8424</issn><eissn>1091-6490</eissn><coden>PNASA6</coden><abstract>Many RNA-associated proteins contain a ribonucleoprotein (RNP) consensus octamer encompassed by a conserved 80 amino acid sequence, which we have termed an RNA recognition motif (RRM). RRM family members contain either one (class I) or multiple (class II) copies of this motif. We report here that a class II component of the U1 small nuclear RNP (snRNP), the A protein of U1 snRNP (U1snRNP-A), contains two RRMs (RRM1 and -2), yet has only one binding domain (RRM1) that interacts specifically with stem-loop II of U1 RNA. Quantitative analysis of binding affinities of fragments of U1snRNP-A demonstrated that an 86-amino acid polypeptide was competent to bind to U1 RNA with an affinity comparable to that of the full-length protein (Kd≈ 80 nM). The carboxyl-terminal RRM2 of U1snRNP-A did not bind to U1 RNA and may recognize an unidentified heterologous RNA. 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| subjects | Amino acids Analytical, structural and metabolic biochemistry Antibodies Binding Sites Biochemistry Biological and medical sciences Electrophoresis, Polyacrylamide Gel Epitopes - analysis Fundamental and applied biological sciences. Psychology Gels Gene Library HeLa cells HeLa Cells - metabolism Humans Kinetics Macromolecular Substances Mutation Nucleic Acid Conformation Nucleic acids Oligonucleotide Probes Protein Biosynthesis Proteins Ribonucleoproteins - genetics Ribonucleoproteins - isolation & purification Ribonucleoproteins - metabolism Ribonucleoproteins, Small Nuclear RNA Rna, ribonucleoproteins RNA, Small Nuclear - metabolism Small nuclear ribonucleoproteins Splicing Transcription, Genetic |
| title | Quantitative Determination that One of Two Potential RNA-Binding Domains of the A Protein Component of the U1 Small Nuclear Ribonucleoprotein Complex Binds with High Affinity to Stem-Loop II of U1 RNA |
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