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Identification Of Small Molecule TRABID Deubiquitinase Inhibitors By Computation-Based Virtual Screen.

Shi T, Bao J, Wang NX, Zheng J, Wu D - BMC Chem Biol (2012)

Bottom Line: However, these inhibitors failed to show inhibitory effects on β-catenin-mediated gene transcription.In addition, expression of TRABID shRNAs, wildtype TRABID, or the DUB activity-deficient mutant showed little effects on β-catenin-mediated gene transcription.TRABID may not be a critical component in canonical Wnt/β-catenin signal transduction or that a minute amount of this protein is sufficient for its role in regulating Wnt activity.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Pharmacology, Yale University School of Medicine, New Haven, CT 06510, USA. jie.zheng@stjude.org.

ABSTRACT

Background: Wnt/β-catenin-mediated gene transcription plays important roles in a wide range of biological and pathophysiological processes including tumorigenesis where β-catenin-mediated transcription activity frequently elevates. TRABID, a deubiquitinase, was shown to have a positive Wnt/β-catenin-mediated gene transcription and hence holds a promise as a putative anti-cancer target.

Results: In this study, we used a combination of structure based virtual screening and an in vitro deubiquitinase (DUB) assay to identify several small molecules that inhibit TRABID DUB activity. However, these inhibitors failed to show inhibitory effects on β-catenin-mediated gene transcription. In addition, expression of TRABID shRNAs, wildtype TRABID, or the DUB activity-deficient mutant showed little effects on β-catenin-mediated gene transcription.

Conclusions: TRABID may not be a critical component in canonical Wnt/β-catenin signal transduction or that a minute amount of this protein is sufficient for its role in regulating Wnt activity.

No MeSH data available.


Related in: MedlinePlus

Effect of TRABID inhibitors on cell growth.  Compounds (20 μM) were added to cells at seeding and cell growth was determined by counting live cells at indicated times. Experiments are performed in triplicates, and data are presented as means ± SD.
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Figure 5: Effect of TRABID inhibitors on cell growth. Compounds (20 μM) were added to cells at seeding and cell growth was determined by counting live cells at indicated times. Experiments are performed in triplicates, and data are presented as means ± SD.

Mentions: Nevertheless, both TRABID inhibitors NSC112200 and NSC267309, but not the non-active analog NSC60650, inhibited the growth of not only the colorectal tumor cell lines HCT-116 and SW480, but also two para-normal cell lines NIH3T3 and HEK293 at 20 μM (Figure 5). These compounds showed no effects on the growth of these cells at 5 μM (data not shown). Because of the lack of effect of Compound NSC60650 on cell growth, we believe that the effects of Compounds NSC112200 and NSC267309 are likely due to their inhibition of the deubiquitinase activity.


Identification Of Small Molecule TRABID Deubiquitinase Inhibitors By Computation-Based Virtual Screen.

Shi T, Bao J, Wang NX, Zheng J, Wu D - BMC Chem Biol (2012)

Effect of TRABID inhibitors on cell growth.  Compounds (20 μM) were added to cells at seeding and cell growth was determined by counting live cells at indicated times. Experiments are performed in triplicates, and data are presented as means ± SD.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3475094&req=5

Figure 5: Effect of TRABID inhibitors on cell growth. Compounds (20 μM) were added to cells at seeding and cell growth was determined by counting live cells at indicated times. Experiments are performed in triplicates, and data are presented as means ± SD.
Mentions: Nevertheless, both TRABID inhibitors NSC112200 and NSC267309, but not the non-active analog NSC60650, inhibited the growth of not only the colorectal tumor cell lines HCT-116 and SW480, but also two para-normal cell lines NIH3T3 and HEK293 at 20 μM (Figure 5). These compounds showed no effects on the growth of these cells at 5 μM (data not shown). Because of the lack of effect of Compound NSC60650 on cell growth, we believe that the effects of Compounds NSC112200 and NSC267309 are likely due to their inhibition of the deubiquitinase activity.

Bottom Line: However, these inhibitors failed to show inhibitory effects on β-catenin-mediated gene transcription.In addition, expression of TRABID shRNAs, wildtype TRABID, or the DUB activity-deficient mutant showed little effects on β-catenin-mediated gene transcription.TRABID may not be a critical component in canonical Wnt/β-catenin signal transduction or that a minute amount of this protein is sufficient for its role in regulating Wnt activity.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Pharmacology, Yale University School of Medicine, New Haven, CT 06510, USA. jie.zheng@stjude.org.

ABSTRACT

Background: Wnt/β-catenin-mediated gene transcription plays important roles in a wide range of biological and pathophysiological processes including tumorigenesis where β-catenin-mediated transcription activity frequently elevates. TRABID, a deubiquitinase, was shown to have a positive Wnt/β-catenin-mediated gene transcription and hence holds a promise as a putative anti-cancer target.

Results: In this study, we used a combination of structure based virtual screening and an in vitro deubiquitinase (DUB) assay to identify several small molecules that inhibit TRABID DUB activity. However, these inhibitors failed to show inhibitory effects on β-catenin-mediated gene transcription. In addition, expression of TRABID shRNAs, wildtype TRABID, or the DUB activity-deficient mutant showed little effects on β-catenin-mediated gene transcription.

Conclusions: TRABID may not be a critical component in canonical Wnt/β-catenin signal transduction or that a minute amount of this protein is sufficient for its role in regulating Wnt activity.

No MeSH data available.


Related in: MedlinePlus