Development of a Bioassay for Continuous Monitoring of TWIST1 Activity

TWIST1 is a transcription factor that affects cell behavior during development and cell differentiation. Yet, it is better known for its roles in neoplasia through regulation of cell plasticity. The pathological contributions of TWIST1 in tumor initiation, angiogenesis, invasion, metastasis, and chemo-resistance have been the focus of much research. To-date, the only way to quantitatively measure the abundance of TWIST is by immunoblots. Yet, no bioassay exists that can detect TWIST1 activity. Thus, we present here a TWIST1 cell-based assay that allows measuring the amount of active TWIST1 non-invasively in living cells. The bioassay was characterized against previously described TWIST1 “inhibitors”, as well as by epigenetic modulators of TWIST1 gene expression. Moreover, we tested multiple cell lines, showing that the level of TWIST1 mRNA resembles that of the bioassay. We show that prostate cancer cells (PC3) undergoing EMT, migrate out of 3D-spheroids and have increased TWIST1 activity. This fast and reliable system to detect active TWIST1 in different biological conditions allows a detailed analysis of this factor, as well as it can be used for drug discovery, since TWIST1 is a potential target for cancer chemotherapeutics.

Download Full-text

Androgen signaling uses a writer and a reader of ADP-ribosylation to regulate protein complex assembly

Nature Communications ◽

10.1038/s41467-021-23055-6 ◽

2021 ◽

Vol 12 (1) ◽

Author(s):

Chun-Song Yang ◽

Kasey Jividen ◽

Teddy Kamata ◽

Natalia Dworak ◽

Luke Oostdyk ◽

...

Keyword(s):

Gene Expression ◽

Protein Interactions ◽

Prostate Cancer Cells ◽

Protein Protein Interactions ◽

Post Translational Modification ◽

Complex Assembly ◽

Adp Ribosylation ◽

Signaling Axis ◽

Regulate Protein ◽

Protein Complex Assembly

AbstractAndrogen signaling through the androgen receptor (AR) directs gene expression in both normal and prostate cancer cells. Androgen regulates multiple aspects of the AR life cycle, including its localization and post-translational modification, but understanding how modifications are read and integrated with AR activity has been difficult. Here, we show that ADP-ribosylation regulates AR through a nuclear pathway mediated by Parp7. We show that Parp7 mono-ADP-ribosylates agonist-bound AR, and that ADP-ribosyl-cysteines within the N-terminal domain mediate recruitment of the E3 ligase Dtx3L/Parp9. Molecular recognition of ADP-ribosyl-cysteine is provided by tandem macrodomains in Parp9, and Dtx3L/Parp9 modulates expression of a subset of AR-regulated genes. Parp7, ADP-ribosylation of AR, and AR-Dtx3L/Parp9 complex assembly are inhibited by Olaparib, a compound used clinically to inhibit poly-ADP-ribosyltransferases Parp1/2. Our study reveals the components of an androgen signaling axis that uses a writer and reader of ADP-ribosylation to regulate protein-protein interactions and AR activity.

Download Full-text

Regulation and imaging of gene expression via an RNA interference antagonistic biomimetic probe

Chemical Science ◽

10.1039/c7sc00909g ◽

2017 ◽

Vol 8 (7) ◽

pp. 4973-4977 ◽

Cited By ~ 12

Author(s):

Kai Zhang ◽

Xue-Jiao Yang ◽

Wei Zhao ◽

Ming-Chen Xu ◽

Jing-Juan Xu ◽

...

Keyword(s):

Gene Expression ◽

Gene Regulation ◽

Rna Interference ◽

Living Cells

A versatile strategy is reported which permits gene regulation and imaging in living cells via an RNA interference antagonistic probe.

Download Full-text

Comprehensive gene expression microarray analysis of Ets-1 blockade in PC3 prostate cancer cells and correlations with prostate cancer tissues: Insights into genes involved in the metastatic cascade

International Journal of Molecular Medicine ◽

10.3892/ijmm.2011.652 ◽

2011 ◽

Vol 27 (6) ◽

Author(s):

Wernert

Keyword(s):

Gene Expression ◽

Prostate Cancer ◽

Cancer Cells ◽

Microarray Analysis ◽

Gene Expression Microarray ◽

Prostate Cancer Cells ◽

Expression Microarray ◽

Metastatic Cascade ◽

Cancer Tissues ◽

Gene Expression Microarray Analysis

Download Full-text

5-Formyluracil targeted biochemical reactions with proteins inhibit DNA replication, induce mutations and interference gene expression in living cells

Chinese Chemical Letters ◽

10.1016/j.cclet.2021.02.036 ◽

2021 ◽

Author(s):

Guangrong Zou ◽

Kaiyuan Zhang ◽

Wei Yang ◽

Chaoxing Liu ◽

Zhentian Fang ◽

...

Keyword(s):

Gene Expression ◽

Dna Replication ◽

Living Cells ◽

Biochemical Reactions

Download Full-text

Regulation of Fertility, Survival, and Cuticle Collagen Function by the Caenorhabditis elegans eaf-1 and ell-1 Genes

Journal of Biological Chemistry ◽

10.1074/jbc.m111.270454 ◽

2011 ◽

Vol 286 (41) ◽

pp. 35915-35921 ◽

Cited By ~ 23

Author(s):

Liquan Cai ◽

Binh L. Phong ◽

Alfred L. Fisher ◽

Zhou Wang

Keyword(s):

Caenorhabditis Elegans ◽

Cell Types ◽

Transcriptional Elongation ◽

Human Prostate Cancer ◽

Prostate Cancer Cells ◽

Transcription Factor Family ◽

Collagen Expression ◽

Extracellular Matrix Components ◽

Multiple Cell

EAF2, an androgen-regulated protein, interacts with members of the ELL (eleven-nineteen lysine-rich leukemia) transcription factor family and also acts as a tumor suppressor. Although these proteins control transcriptional elongation and perhaps modulate the effects of other transcription factors, the mechanisms of their actions remain largely unknown. To gain new insights into the biology of the EAF2 and ELL family proteins, we used Caenorhabditis elegans as a model to explore the in vivo roles of their worm orthologs. Through the use of transgenic worms, RNAi, and an eaf-1 mutant, we found that both genes are expressed in multiple cell types throughout the worm life cycle and that they play important roles in fertility, survival, and body size regulation. ELL-1 and EAF-1 likely contribute to these activities in part through modulating cuticle synthesis, given that we observed a disrupted cuticle structure in ell-1 RNAi-treated or eaf-1 mutant worms. Consistent with disruption of cuticle structure, loss of either ELL-1 or EAF-1 suppressed the rol phenotype of specific collagen mutants, possibly through the control of dpy-3, dpy-13, and sqt-3 collagen gene expression. Furthermore, we also noted the regulation of collagen expression by ELL overexpression in PC3 human prostate cancer cells. Together, these results reveal important roles for the eaf-1 and ell-1 genes in the regulation of extracellular matrix components.

Download Full-text