Chapter title |
RNA-Protein Complexes and Interactions
|
---|---|
Chapter number | 15 |
Book title |
RNA-Protein Complexes and Interactions
|
Published in |
Methods in molecular biology, January 2016
|
DOI | 10.1007/978-1-4939-3591-8_15 |
Pubmed ID | |
Book ISBNs |
978-1-4939-3589-5, 978-1-4939-3591-8
|
Authors |
Yeh, Fu-Lung, Tung, Luh, Chang, Tien-Hsien, Fu-Lung Yeh, Luh Tung, Tien-Hsien Chang |
Abstract |
Although DExD/H-box proteins are known to unwind RNA duplexes and modulate RNA structures in vitro, it is highly plausible that, in vivo, some may function to remodel RNA-protein complexes. Precisely how the latter is achieved remains a mystery. We investigated this critical issue by using yeast Prp28p, an evolutionarily conserved DExD/H-box splicing factor, as a model system. To probe how Prp28p interacts with spliceosome, we strategically placed p-benzoyl-phenylalanine (BPA), a photoactivatable unnatural amino acid, along the body of Prp28p in vivo. Extracts prepared from these engineered strains were then used to assemble in vitro splicing reactions for BPA-mediated protein-protein crosslinkings. This enabled us, for the first time, to "capture" Prp28p in action. This approach may be applicable to studying the roles of other DExD/H-box proteins functioning in diverse RNA-related pathways, as well as to investigating protein-protein contacts within an RNA-protein complex. |
X Demographics
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 1 | 100% |
Demographic breakdown
Type | Count | As % |
---|---|---|
Members of the public | 1 | 100% |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 5 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Student > Ph. D. Student | 2 | 40% |
Researcher | 2 | 40% |
Unknown | 1 | 20% |
Readers by discipline | Count | As % |
---|---|---|
Biochemistry, Genetics and Molecular Biology | 1 | 20% |
Agricultural and Biological Sciences | 1 | 20% |
Neuroscience | 1 | 20% |
Medicine and Dentistry | 1 | 20% |
Unknown | 1 | 20% |