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Small angle X-ray scattering studies of CTNNBL1 dimerization and CTNNBL1/CDC5L complex
The hPrp19/CDC5L complex is a non-snRNP spliceosome complex that plays a key role in the spliceosome activation during pre-mRNA splicing, and CTNNBL1 and CDC5L are essential components of the complex. In this study, to investigate the oligomeric state of CTNNBL1 in solution, we performed small angle...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4585563/ https://www.ncbi.nlm.nih.gov/pubmed/26381213 http://dx.doi.org/10.1038/srep14251 |
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author | Ahn, Jae-Woo Sik Jin, Kyeong Francis Son, Hyeoncheol Ho Chang, Jeong Kim, Kyung-Jin |
author_facet | Ahn, Jae-Woo Sik Jin, Kyeong Francis Son, Hyeoncheol Ho Chang, Jeong Kim, Kyung-Jin |
author_sort | Ahn, Jae-Woo |
collection | PubMed |
description | The hPrp19/CDC5L complex is a non-snRNP spliceosome complex that plays a key role in the spliceosome activation during pre-mRNA splicing, and CTNNBL1 and CDC5L are essential components of the complex. In this study, to investigate the oligomeric state of CTNNBL1 in solution, we performed small angle X-ray scattering experiments in various concentrations of NaCl. We observed that CTNNBL1 existed as a dimer in physiological NaCl concentrations. Site-directed mutagenesis experiment of CTNNBL1 confirmed that N-terminal capping region and the first four ARM repeats are important for dimerization of the protein. We also found that the positively-charged NLS3-containing region (residues 197–235) of CDC5L bound to the negatively-charged patch of CTNNBL1 and that the CTNNBL1/CDC5L complex formed a heterotetramer consisting of one CTNNBL1 dimer and one CDC5L dimer. Moreover, reconstruction of 3D models of CTNNBL1/CDC5L complexes containing CTNNBL1 and three different truncated forms of CDC5L showed that the CDC5L(141–196) region and the CDC5L(236–377) region were positioned at the top of the N-terminal capping region and at the bottom of ARM VII of CTNNBL1, respectively. |
format | Online Article Text |
id | pubmed-4585563 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | Nature Publishing Group |
record_format | MEDLINE/PubMed |
spelling | pubmed-45855632015-09-29 Small angle X-ray scattering studies of CTNNBL1 dimerization and CTNNBL1/CDC5L complex Ahn, Jae-Woo Sik Jin, Kyeong Francis Son, Hyeoncheol Ho Chang, Jeong Kim, Kyung-Jin Sci Rep Article The hPrp19/CDC5L complex is a non-snRNP spliceosome complex that plays a key role in the spliceosome activation during pre-mRNA splicing, and CTNNBL1 and CDC5L are essential components of the complex. In this study, to investigate the oligomeric state of CTNNBL1 in solution, we performed small angle X-ray scattering experiments in various concentrations of NaCl. We observed that CTNNBL1 existed as a dimer in physiological NaCl concentrations. Site-directed mutagenesis experiment of CTNNBL1 confirmed that N-terminal capping region and the first four ARM repeats are important for dimerization of the protein. We also found that the positively-charged NLS3-containing region (residues 197–235) of CDC5L bound to the negatively-charged patch of CTNNBL1 and that the CTNNBL1/CDC5L complex formed a heterotetramer consisting of one CTNNBL1 dimer and one CDC5L dimer. Moreover, reconstruction of 3D models of CTNNBL1/CDC5L complexes containing CTNNBL1 and three different truncated forms of CDC5L showed that the CDC5L(141–196) region and the CDC5L(236–377) region were positioned at the top of the N-terminal capping region and at the bottom of ARM VII of CTNNBL1, respectively. Nature Publishing Group 2015-09-18 /pmc/articles/PMC4585563/ /pubmed/26381213 http://dx.doi.org/10.1038/srep14251 Text en Copyright © 2015, Macmillan Publishers Limited http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ |
spellingShingle | Article Ahn, Jae-Woo Sik Jin, Kyeong Francis Son, Hyeoncheol Ho Chang, Jeong Kim, Kyung-Jin Small angle X-ray scattering studies of CTNNBL1 dimerization and CTNNBL1/CDC5L complex |
title | Small angle X-ray scattering studies of CTNNBL1 dimerization and CTNNBL1/CDC5L complex |
title_full | Small angle X-ray scattering studies of CTNNBL1 dimerization and CTNNBL1/CDC5L complex |
title_fullStr | Small angle X-ray scattering studies of CTNNBL1 dimerization and CTNNBL1/CDC5L complex |
title_full_unstemmed | Small angle X-ray scattering studies of CTNNBL1 dimerization and CTNNBL1/CDC5L complex |
title_short | Small angle X-ray scattering studies of CTNNBL1 dimerization and CTNNBL1/CDC5L complex |
title_sort | small angle x-ray scattering studies of ctnnbl1 dimerization and ctnnbl1/cdc5l complex |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4585563/ https://www.ncbi.nlm.nih.gov/pubmed/26381213 http://dx.doi.org/10.1038/srep14251 |
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