Cytoplasmic, full length and novel cleaved variant, TBLR1 reduces apoptosis in prostate cancer under androgen deprivation

Garrett Daniels, Xinmin Zhang, Xuelin Zhong, Larion Santiago, Ling Hang Wang, Xinyu Wu, Jack Y. Zhang, Fengxia Liang, Xin Li, Thomas A. Neubert, Laurey Steinke, Ying Shen, Ross Basch, Robert Schneider, David E. Levy, Peng Lee

Research output: Contribution to journalArticle

Abstract

TBLR1/TBL1XR1, a core component of the nuclear receptor corepressor (NCoR) complex critical for the regulation of multiple nuclear receptors, is a transcriptional coactivator of androgen receptor (AR) and functions as a tumor suppressor when expressed in the nucleus in prostate. Subcellular localization of a protein is critical for its function, and although TBLR1, as a transcriptional cofactor, has been primarily viewed as a nuclear protein, many cells also express variable levels of cytoplasmic TBLR1 and its cytoplasmic specific functions have not been studied. Prostate cancer (PCa) cells express moderately higher level of cytoplasmic TBLR1 compared to benign prostate cells. When comparing androgen-dependent (AD) to androgen-independent (AI) PCa, AI cells contain very high levels of TBLR1 cytoplasmic expression and low levels of nuclear expression. Overexpression of cytoplasmic TBLR1 in AD cells inhibits apoptosis induced by androgen deprivation therapy, either in an androgen free condition or in the presence of bicalutamide. Additionally, we identified a cytoplasmic specific isoform of TBLR1 (cvTBLR1) approximately 5 kDa lower in molecular weight, that is expressed at higher levels in AI PCa cells. By immunoprecipitation, we purified cvTBLR1 and using mass spectrometry analysis combined with N-terminal TMPP labeling and Edman degradation, we identified the cleavage site of cvTBLR1 at amino acid 89, truncating the first 88 amino acids of the N-terminus of the full length protein. Functionally, cvTBLR1 expressed in the cytoplasm reduced apoptosis in PCa cells and promoted growth, migration, and invasion. Finally, we identified a nuclear export signal sequence for TBLR1 cellular localization by deletion and sitedirected mutagenesis. The roles of TBLR1 and cvTBLR1 provide novel insights into the mechanism of castration resistance and new strategies for PCa therapy.

Original languageEnglish (US)
Pages (from-to)39556-39571
Number of pages16
JournalOncotarget
Volume7
Issue number26
StatePublished - 2016

Fingerprint

Androgens
Prostatic Neoplasms
Apoptosis
Prostate
Nuclear Export Signals
Amino Acids
Co-Repressor Proteins
Castration
Androgen Receptors
Cytoplasmic and Nuclear Receptors
Nuclear Proteins
Protein Sorting Signals
Immunoprecipitation
Mutagenesis
Mass Spectrometry
Protein Isoforms
Cytoplasm
Proteins
Molecular Weight
Therapeutics

Keywords

  • Castration resistance
  • cvTBLR1
  • Prostate cancer
  • Subcellular localization
  • TBLR1

ASJC Scopus subject areas

  • Oncology

Cite this

Daniels, G., Zhang, X., Zhong, X., Santiago, L., Wang, L. H., Wu, X., ... Lee, P. (2016). Cytoplasmic, full length and novel cleaved variant, TBLR1 reduces apoptosis in prostate cancer under androgen deprivation. Oncotarget, 7(26), 39556-39571.

Cytoplasmic, full length and novel cleaved variant, TBLR1 reduces apoptosis in prostate cancer under androgen deprivation. / Daniels, Garrett; Zhang, Xinmin; Zhong, Xuelin; Santiago, Larion; Wang, Ling Hang; Wu, Xinyu; Zhang, Jack Y.; Liang, Fengxia; Li, Xin; Neubert, Thomas A.; Steinke, Laurey; Shen, Ying; Basch, Ross; Schneider, Robert; Levy, David E.; Lee, Peng.

In: Oncotarget, Vol. 7, No. 26, 2016, p. 39556-39571.

Research output: Contribution to journalArticle

Daniels, G, Zhang, X, Zhong, X, Santiago, L, Wang, LH, Wu, X, Zhang, JY, Liang, F, Li, X, Neubert, TA, Steinke, L, Shen, Y, Basch, R, Schneider, R, Levy, DE & Lee, P 2016, 'Cytoplasmic, full length and novel cleaved variant, TBLR1 reduces apoptosis in prostate cancer under androgen deprivation', Oncotarget, vol. 7, no. 26, pp. 39556-39571.
Daniels G, Zhang X, Zhong X, Santiago L, Wang LH, Wu X et al. Cytoplasmic, full length and novel cleaved variant, TBLR1 reduces apoptosis in prostate cancer under androgen deprivation. Oncotarget. 2016;7(26):39556-39571.
Daniels, Garrett ; Zhang, Xinmin ; Zhong, Xuelin ; Santiago, Larion ; Wang, Ling Hang ; Wu, Xinyu ; Zhang, Jack Y. ; Liang, Fengxia ; Li, Xin ; Neubert, Thomas A. ; Steinke, Laurey ; Shen, Ying ; Basch, Ross ; Schneider, Robert ; Levy, David E. ; Lee, Peng. / Cytoplasmic, full length and novel cleaved variant, TBLR1 reduces apoptosis in prostate cancer under androgen deprivation. In: Oncotarget. 2016 ; Vol. 7, No. 26. pp. 39556-39571.
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abstract = "TBLR1/TBL1XR1, a core component of the nuclear receptor corepressor (NCoR) complex critical for the regulation of multiple nuclear receptors, is a transcriptional coactivator of androgen receptor (AR) and functions as a tumor suppressor when expressed in the nucleus in prostate. Subcellular localization of a protein is critical for its function, and although TBLR1, as a transcriptional cofactor, has been primarily viewed as a nuclear protein, many cells also express variable levels of cytoplasmic TBLR1 and its cytoplasmic specific functions have not been studied. Prostate cancer (PCa) cells express moderately higher level of cytoplasmic TBLR1 compared to benign prostate cells. When comparing androgen-dependent (AD) to androgen-independent (AI) PCa, AI cells contain very high levels of TBLR1 cytoplasmic expression and low levels of nuclear expression. Overexpression of cytoplasmic TBLR1 in AD cells inhibits apoptosis induced by androgen deprivation therapy, either in an androgen free condition or in the presence of bicalutamide. Additionally, we identified a cytoplasmic specific isoform of TBLR1 (cvTBLR1) approximately 5 kDa lower in molecular weight, that is expressed at higher levels in AI PCa cells. By immunoprecipitation, we purified cvTBLR1 and using mass spectrometry analysis combined with N-terminal TMPP labeling and Edman degradation, we identified the cleavage site of cvTBLR1 at amino acid 89, truncating the first 88 amino acids of the N-terminus of the full length protein. Functionally, cvTBLR1 expressed in the cytoplasm reduced apoptosis in PCa cells and promoted growth, migration, and invasion. Finally, we identified a nuclear export signal sequence for TBLR1 cellular localization by deletion and sitedirected mutagenesis. The roles of TBLR1 and cvTBLR1 provide novel insights into the mechanism of castration resistance and new strategies for PCa therapy.",
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AU - Zhang, Jack Y.

AU - Liang, Fengxia

AU - Li, Xin

AU - Neubert, Thomas A.

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