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Structure-Function Studies of the bHLH Phosphorylation Domain of TWIST1 in Prostate Cancer Cells()()
The TWIST1 gene has diverse roles in development and pathologic diseases such as cancer. TWIST1 is a dimeric basic helix-loop-helix (bHLH) transcription factor existing as TWIST1-TWIST1 or TWIST1-E12/47. TWIST1 partner choice and DNA binding can be influenced during development by phosphorylation of...
Autores principales: | , , , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Neoplasia Press
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4309734/ https://www.ncbi.nlm.nih.gov/pubmed/25622896 http://dx.doi.org/10.1016/j.neo.2014.10.009 |
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author | Gajula, Rajendra P. Chettiar, Sivarajan T. Williams, Russell D. Nugent, Katriana Kato, Yoshinori Wang, Hailun Malek, Reem Taparra, Kekoa Cades, Jessica Annadanam, Anvesh Yoon, A-Rum Fertig, Elana Firulli, Beth A. Mazzacurati, Lucia Burns, Timothy F. Firulli, Anthony B. An, Steven S. Tran, Phuoc T. |
author_facet | Gajula, Rajendra P. Chettiar, Sivarajan T. Williams, Russell D. Nugent, Katriana Kato, Yoshinori Wang, Hailun Malek, Reem Taparra, Kekoa Cades, Jessica Annadanam, Anvesh Yoon, A-Rum Fertig, Elana Firulli, Beth A. Mazzacurati, Lucia Burns, Timothy F. Firulli, Anthony B. An, Steven S. Tran, Phuoc T. |
author_sort | Gajula, Rajendra P. |
collection | PubMed |
description | The TWIST1 gene has diverse roles in development and pathologic diseases such as cancer. TWIST1 is a dimeric basic helix-loop-helix (bHLH) transcription factor existing as TWIST1-TWIST1 or TWIST1-E12/47. TWIST1 partner choice and DNA binding can be influenced during development by phosphorylation of Thr125 and Ser127 of the Thr-Gln-Ser (TQS) motif within the bHLH of TWIST1. The significance of these TWIST1 phosphorylation sites for metastasis is unknown. We created stable isogenic prostate cancer cell lines overexpressing TWIST1 wild-type, phospho-mutants, and tethered versions. We assessed these isogenic lines using assays that mimic stages of cancer metastasis. In vitro assays suggested the phospho-mimetic Twist1-DQD mutation could confer cellular properties associated with pro-metastatic behavior. The hypo-phosphorylation mimic Twist1-AQA mutation displayed reduced pro-metastatic activity compared to wild-type TWIST1 in vitro, suggesting that phosphorylation of the TWIST1 TQS motif was necessary for pro-metastatic functions. In vivo analysis demonstrates that the Twist1-AQA mutation exhibits reduced capacity to contribute to metastasis, whereas the expression of the Twist1-DQD mutation exhibits proficient metastatic potential. Tethered TWIST1-E12 heterodimers phenocopied the Twist1-DQD mutation for many in vitro assays, suggesting that TWIST1 phosphorylation may result in heterodimerization in prostate cancer cells. Lastly, the dual phosphatidylinositide 3-kinase (PI3K)-mammalian target of rapamycin (mTOR) inhibitor BEZ235 strongly attenuated TWIST1-induced migration that was dependent on the TQS motif. TWIST1 TQS phosphorylation state determines the intensity of TWIST1-induced pro-metastatic ability in prostate cancer cells, which may be partly explained mechanistically by TWIST1 dimeric partner choice. |
format | Online Article Text |
id | pubmed-4309734 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | Neoplasia Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-43097342015-01-30 Structure-Function Studies of the bHLH Phosphorylation Domain of TWIST1 in Prostate Cancer Cells()() Gajula, Rajendra P. Chettiar, Sivarajan T. Williams, Russell D. Nugent, Katriana Kato, Yoshinori Wang, Hailun Malek, Reem Taparra, Kekoa Cades, Jessica Annadanam, Anvesh Yoon, A-Rum Fertig, Elana Firulli, Beth A. Mazzacurati, Lucia Burns, Timothy F. Firulli, Anthony B. An, Steven S. Tran, Phuoc T. Neoplasia Article The TWIST1 gene has diverse roles in development and pathologic diseases such as cancer. TWIST1 is a dimeric basic helix-loop-helix (bHLH) transcription factor existing as TWIST1-TWIST1 or TWIST1-E12/47. TWIST1 partner choice and DNA binding can be influenced during development by phosphorylation of Thr125 and Ser127 of the Thr-Gln-Ser (TQS) motif within the bHLH of TWIST1. The significance of these TWIST1 phosphorylation sites for metastasis is unknown. We created stable isogenic prostate cancer cell lines overexpressing TWIST1 wild-type, phospho-mutants, and tethered versions. We assessed these isogenic lines using assays that mimic stages of cancer metastasis. In vitro assays suggested the phospho-mimetic Twist1-DQD mutation could confer cellular properties associated with pro-metastatic behavior. The hypo-phosphorylation mimic Twist1-AQA mutation displayed reduced pro-metastatic activity compared to wild-type TWIST1 in vitro, suggesting that phosphorylation of the TWIST1 TQS motif was necessary for pro-metastatic functions. In vivo analysis demonstrates that the Twist1-AQA mutation exhibits reduced capacity to contribute to metastasis, whereas the expression of the Twist1-DQD mutation exhibits proficient metastatic potential. Tethered TWIST1-E12 heterodimers phenocopied the Twist1-DQD mutation for many in vitro assays, suggesting that TWIST1 phosphorylation may result in heterodimerization in prostate cancer cells. Lastly, the dual phosphatidylinositide 3-kinase (PI3K)-mammalian target of rapamycin (mTOR) inhibitor BEZ235 strongly attenuated TWIST1-induced migration that was dependent on the TQS motif. TWIST1 TQS phosphorylation state determines the intensity of TWIST1-induced pro-metastatic ability in prostate cancer cells, which may be partly explained mechanistically by TWIST1 dimeric partner choice. Neoplasia Press 2015-01-23 /pmc/articles/PMC4309734/ /pubmed/25622896 http://dx.doi.org/10.1016/j.neo.2014.10.009 Text en © 2014Neoplasia Press, Inc. Published by Elsevier Inc. http://creativecommons.org/licenses/by-nc-nd/3.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/3.0/). |
spellingShingle | Article Gajula, Rajendra P. Chettiar, Sivarajan T. Williams, Russell D. Nugent, Katriana Kato, Yoshinori Wang, Hailun Malek, Reem Taparra, Kekoa Cades, Jessica Annadanam, Anvesh Yoon, A-Rum Fertig, Elana Firulli, Beth A. Mazzacurati, Lucia Burns, Timothy F. Firulli, Anthony B. An, Steven S. Tran, Phuoc T. Structure-Function Studies of the bHLH Phosphorylation Domain of TWIST1 in Prostate Cancer Cells()() |
title | Structure-Function Studies of the bHLH Phosphorylation Domain of TWIST1 in Prostate Cancer Cells()() |
title_full | Structure-Function Studies of the bHLH Phosphorylation Domain of TWIST1 in Prostate Cancer Cells()() |
title_fullStr | Structure-Function Studies of the bHLH Phosphorylation Domain of TWIST1 in Prostate Cancer Cells()() |
title_full_unstemmed | Structure-Function Studies of the bHLH Phosphorylation Domain of TWIST1 in Prostate Cancer Cells()() |
title_short | Structure-Function Studies of the bHLH Phosphorylation Domain of TWIST1 in Prostate Cancer Cells()() |
title_sort | structure-function studies of the bhlh phosphorylation domain of twist1 in prostate cancer cells()() |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4309734/ https://www.ncbi.nlm.nih.gov/pubmed/25622896 http://dx.doi.org/10.1016/j.neo.2014.10.009 |
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