Cargando…

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...

Descripción completa

Detalles Bibliográficos
Autores principales: 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.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Neoplasia Press 2015
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
_version_ 1782354744435539968
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
work_keys_str_mv AT gajularajendrap structurefunctionstudiesofthebhlhphosphorylationdomainoftwist1inprostatecancercells
AT chettiarsivarajant structurefunctionstudiesofthebhlhphosphorylationdomainoftwist1inprostatecancercells
AT williamsrusselld structurefunctionstudiesofthebhlhphosphorylationdomainoftwist1inprostatecancercells
AT nugentkatriana structurefunctionstudiesofthebhlhphosphorylationdomainoftwist1inprostatecancercells
AT katoyoshinori structurefunctionstudiesofthebhlhphosphorylationdomainoftwist1inprostatecancercells
AT wanghailun structurefunctionstudiesofthebhlhphosphorylationdomainoftwist1inprostatecancercells
AT malekreem structurefunctionstudiesofthebhlhphosphorylationdomainoftwist1inprostatecancercells
AT taparrakekoa structurefunctionstudiesofthebhlhphosphorylationdomainoftwist1inprostatecancercells
AT cadesjessica structurefunctionstudiesofthebhlhphosphorylationdomainoftwist1inprostatecancercells
AT annadanamanvesh structurefunctionstudiesofthebhlhphosphorylationdomainoftwist1inprostatecancercells
AT yoonarum structurefunctionstudiesofthebhlhphosphorylationdomainoftwist1inprostatecancercells
AT fertigelana structurefunctionstudiesofthebhlhphosphorylationdomainoftwist1inprostatecancercells
AT firullibetha structurefunctionstudiesofthebhlhphosphorylationdomainoftwist1inprostatecancercells
AT mazzacuratilucia structurefunctionstudiesofthebhlhphosphorylationdomainoftwist1inprostatecancercells
AT burnstimothyf structurefunctionstudiesofthebhlhphosphorylationdomainoftwist1inprostatecancercells
AT firullianthonyb structurefunctionstudiesofthebhlhphosphorylationdomainoftwist1inprostatecancercells
AT anstevens structurefunctionstudiesofthebhlhphosphorylationdomainoftwist1inprostatecancercells
AT tranphuoct structurefunctionstudiesofthebhlhphosphorylationdomainoftwist1inprostatecancercells