Project description:Deubiquitinases (DUBs) play a role in regulating protein degradation which is critical for maintaining protein homeostasis. The purpose of this study was to verify biological consequence of genetic depletion of USP21 in human cells on proteome changes and cellular function. Comparison of proteomic analysis of HAP-1 USP21 knockout and HAP-1 WT cells revealed that down-regulated proteins in HAP-1 USP21 KO cell are mostly engaged in mitochondrial function and ATP production. Cellular assays confirmed disturbance of mitochondrial metabolism and decrease in ATP production in HAP-1 USP21 knock-out cells.
Project description:Analysis of CPEB translational regulator target mRNAs Microarray analysis of mRNAs associated with polysomes in wild type (WT) and Cpeb1 KO MEFs
Project description:This first-in-human (FIH) dose-escalation and dose-validation/expansion study will assess KO-2806, a farnesyl transferase inhibitor (FTI), as a monotherapy and in combination, in adult patients with advanced solid tumors.
Project description:liquid chromatographytandem-mass spectrometry (LC-MS) and proteomic analysis were conducted to analyze the differential expression proteins (DEPs) in WT and SCRN1 KO cells
Project description:Activating Transcription Factor 4 (ATF4) is a transcription factor induced by the integrated stress response (ISR). This experiment is a genome-wide profiling of ATF4-dependent RNA expression in human HAP-1 cells. HAP-1 is a near-haploid human cell line that was derived from KBM-7 cells isolated from a patient with Chronic Myelogenous Leukemia. We analyzed WT and ATF4 KO cells. We induced ATF4 expression by mimicking amino acid starvation with the drug histidinol. RNA expression profiles were generated for WT and ATF4 KO HAP1 cells. ATF4 genes were mutated using Cas9 genome editing technology. Amino acid starvation was mimicked by treating WT and ATF4 KO cells with 2 mM histidinol for 24 hours, which increases ATF4 expression.
Project description:Cytokinesis is a key step in the later stage of cell division to partition cellular contents into daughter cells, which undergoes precise regulation in temporal and spatial. However, the underlying mechanisms are still unclear. Here, we show that Hsp90 cochaperone, NudCL2 (NudC-like protein 2), is required for cytokinesis in mammalian cells. NudCL2 localizes to midbody during cytokinesis. Knockout (KO) of NudCL2 using CRISPR/Cas9-based genome editing causes cytokinesis failure and leads to the accumulation of multinucleated cells. To investigate the potential regulator involved in NudCL2-mediated cytokinesis regulation, we performed isobaric tags for relative and absolute quantitation (iTRAQ)-based quantitative proteomic analysis in WT and NudCL2 KO cells and found hundreds of differentially expressed proteins (KO/WT fold change > 1.2 or < 0.83, p < 0.05) in NudCL2 KO cells. We found that the protein level of RCC2 (a midbody-associated protein) is obviously decrease in NudCL2 KO cells. Further studies display that knockout of NudCL2 induces an increase in RCC2 protein instability and degradation. Our data show that depletion of RCC2 leads to the similar defects as NudCL2 KO. Ectopic expression of RCC2 effectively rescues the cytokinesis failure caused by the loss of NudCL2. Interestingly, our data reveals that Hsp90 localizes to midbody in cytokinesis, and interacts with NudCL2 and RCC2. Inhibition of Hsp90 ATPase activity also causes the RCC2 instability and cytokinesis failure as NudCL2 KO. Moreover, the abnormal phenotypes induced by NudCL2 KO are able to rescue by the ectopic expression of Hsp90, but not vice versa. Taken together, our data suggest that NudCL2 is required to cytokinesis by stabilizing RCC2 at midbody through the Hsp90 pathway, providing a hitherto unrecognized mechanism crucial for cytokinesis regulation.
Project description:mRNAseq and proteomic data set of one week old WT (Chop wt/wt CkmmCre wt/wt Dars2 fl/fl), Chop KO (Chop ko/ko CkmmCre wt/wt Dars2 fl/fl), Dars2 KO (Chop wt/wt CkmmCre tg/wt Dars2 fl/fl) and DKO (Chop ko/ko CkmmCre tg/wt Dars2 fl/fl) mice