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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
25
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pubmed:dateCreated |
1996-1-24
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pubmed:abstractText |
An in vitro genetic system was developed as a rapid means for studying the specificity determinants of RNA-binding proteins. This system was used to investigate the origin of the RNA-binding specificity of the mammalian spliceosomal protein U1A. The U1A domain responsible for binding to U1 small nuclear RNA was locally mutagenized and displayed as a combinatorial library on filamentous bacteriophage. Affinity selection identified four U1A residues in the mutagenized region that are important for specific binding to U1 hairpin II. One of these residues (Leu-49) disproportionately affects the rates of binding and release and appears to play a critical role in locking the protein onto the RNA. Interestingly, a protein variant that binds more tightly than U1A emerged during the selection, showing that the affinity of U1A for U1 RNA has not been optimized during evolution.
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pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/8524863-1826042,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8524863-1826055,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8524863-1832492,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8524863-1833186,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8524863-1908075,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8524863-1934059,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8524863-2140872,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8524863-2147232,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8524863-2199796,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8524863-2200121,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8524863-4001944,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8524863-7520277,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8524863-7984237,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8524863-8036511,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8524863-8262062,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8524863-8500177
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
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pubmed:status |
MEDLINE
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pubmed:month |
Dec
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pubmed:issn |
0027-8424
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:day |
5
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pubmed:volume |
92
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
11859-63
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pubmed:dateRevised |
2009-11-19
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pubmed:meshHeading |
pubmed-meshheading:8524863-Amino Acid Sequence,
pubmed-meshheading:8524863-Bacteriophages,
pubmed-meshheading:8524863-Base Sequence,
pubmed-meshheading:8524863-Binding Sites,
pubmed-meshheading:8524863-DNA, Recombinant,
pubmed-meshheading:8524863-Gene Library,
pubmed-meshheading:8524863-Genetic Variation,
pubmed-meshheading:8524863-Genetic Vectors,
pubmed-meshheading:8524863-Molecular Sequence Data,
pubmed-meshheading:8524863-Nucleic Acid Conformation,
pubmed-meshheading:8524863-RNA, Small Nuclear,
pubmed-meshheading:8524863-RNA-Binding Proteins,
pubmed-meshheading:8524863-Recombinant Proteins,
pubmed-meshheading:8524863-Ribonucleoprotein, U1 Small Nuclear,
pubmed-meshheading:8524863-Selection, Genetic,
pubmed-meshheading:8524863-Structure-Activity Relationship
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pubmed:year |
1995
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pubmed:articleTitle |
Analysis of RNA-binding proteins by in vitro genetic selection: identification of an amino acid residue important for locking U1A onto its RNA target.
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pubmed:affiliation |
Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, MA 02115, USA.
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pubmed:publicationType |
Journal Article,
Comparative Study,
Research Support, Non-U.S. Gov't
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