| Autor: |
Laird-Offringa, I A, Belasco, J G |
| Zdroj: |
Proceedings of the National Academy of Sciences of the United States of America; December 1995, Vol. 92 Issue: 25 p11859-11863, 5p |
| Abstrakt: |
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. |
| Databáze: |
Supplemental Index |
| Externí odkaz: |
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