Identification of Decrease in TRiC Proteins as Novel Targets of Alpha-Amanitin-Derived Hepatotoxicity by Comparative Proteomic Analysis In Vitro

Alpha-amanitin (α-AMA) is a cyclic peptide and one of the most lethal mushroom amatoxins found in Amanita phalloides. α-AMA is known to cause hepatotoxicity through RNA polymerase II inhibition, which acts in RNA and DNA translocation. To investigate the toxic signature of α-AMA beyond known mechanisms, we used quantitative nanoflow liquid chromatography–tandem mass spectrometry analysis coupled with tandem mass tag labeling to examine proteome dynamics in Huh-7 human hepatoma cells treated with toxic concentrations of α-AMA. Among the 1828 proteins identified, we quantified 1563 proteins, which revealed that four subunits in the T-complex protein 1-ring complex protein decreased depending on the α-AMA concentration. We conducted bioinformatics analyses of the quantified proteins to characterize the toxic signature of α-AMA in hepatoma cells. This is the first report of global changes in proteome abundance with variations in α-AMA concentration, and our findings suggest a novel molecular regulation mechanism for hepatotoxicity.

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Faculty Opinions recommendation of Characterisation of the gene encoding a protective antigen from Babesia microti identified it as eta subunit of chaperonin containing T-complex protein 1.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.1002445.27154 ◽

2001 ◽

Author(s):

David Persing

Keyword(s):

Protective Antigen ◽

Babesia Microti ◽

Gene Encoding ◽

T Complex ◽

Complex Protein

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Faculty Opinions recommendation of Structural basis of transcription: alpha-amanitin-RNA polymerase II cocrystal at 2.8 A resolution.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.1004788.55306 ◽

2002 ◽

Author(s):

Ronald Conaway

Keyword(s):

Rna Polymerase ◽

Rna Polymerase Ii ◽

Structural Basis ◽

Alpha Amanitin

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Faculty Opinions recommendation of Structural basis of transcription inhibition by alpha-amanitin and implications for RNA polymerase II translocation.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.1116743.572814 ◽

2008 ◽

Author(s):

Katsuhiko Murakami

Keyword(s):

Rna Polymerase ◽

Rna Polymerase Ii ◽

Structural Basis ◽

Transcription Inhibition ◽

Alpha Amanitin

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The KLF6 Super Enhancer Modulates Cell Proliferation via MiR-1301 in Human Hepatoma Cells

MicroRNA ◽

10.2174/2211536608666190314122725 ◽

2019 ◽

Vol 9 (1) ◽

pp. 64-69 ◽

Cited By ~ 1

Author(s):

KumChol Ri ◽

Chol Kim ◽

CholJin Pak ◽

PhyongChol Ri ◽

HyonChol Om

Keyword(s):

Cell Proliferation ◽

Cellular Proliferation ◽

Hepatoma Cells ◽

Regulation Mechanism ◽

Dependent Manner ◽

Human Hepatoma ◽

Human Hepatoma Cells ◽

Over Expression ◽

Super Enhancer ◽

Engineering Changes

Background: Recent studies have attempted to elucidate the function of super enhancers by means of microRNAs. Although the functional outcomes of miR-1301 have become clearer, the pathways that regulate the expressions of miR-1301 remain unclear. Objective: The objective of this paper was to consider the pathway regulating expression of miR- 1301 and miR-1301 signaling pathways with the inhibition of cell proliferation. Methods: In this study, we prepared the cell clones that the KLF6 super enhancer was deleted by means of the CRISPR/Cas9 system-mediated genetic engineering. Changes in miR-1301 expression after the deletion of the KLF6 super enhancer were evaluated by RT-PCR analysis, and the signal pathway of miR-1301 with inhibition of the cell proliferation was examined using RNA interference technology. Results: The results showed that miR-1301 expression was significantly increased after the deletion of the KLF6 super enhancer. Over-expression of miR-1301 induced by deletion of the KLF6 super enhancer also regulated the expression of p21 and p53 in human hepatoma cells. functional modeling of findings using siRNA specific to miR-1301 showed that expression level changes had direct biological effects on cellular proliferation in Human hepatoma cells. Furthermore, cellular proliferation assay was shown to be directly associated with miR-1301 levels. Conclusion: As a result, it was demonstrated that the over-expression of miR-1301 induced by the disruption of the KLF6 super enhancer leads to a significant inhibition of proliferation in HepG2 cells. Moreover, it was demonstrated that the KLF6 super enhancer regulates the cell-proliferative effects which are mediated, at least in part, by the induction of p21and p53 in a p53-dependent manner. Our results provide the functional significance of miR-1301 in understanding the transcriptional regulation mechanism of the KLF6 super enhancer.

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