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OTUD6A increases the amount of CDC6. 12 / 12
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"Depletion of OTUD6A inhibits CDC6 expression as well as BBN-induced BCa tumorigenesis and tumour progression."
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"Collectively, these results demonstrate that OTUD6A positively regulates the CDC6 protein level both in vivo and in vitro."
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"Immunofluorescence (IF) staining further confirmed that overexpression of the OTUD6A upregulated the endogenous CDC6 protein level, compared to those transfected with empty Flag vector and Flag-OTUD6A untransfected cells (Fig. 1d), and transient transfection with empty Flag vector or Flag-OTUD6A plasmid didn’t change the cell cycle distribution (Supplementary Fig. 1b)."
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"Thus we fractionated cell lysates into cytoplasmic, soluble and chromatin-bound fractions, and found that overexpression of OTUD6A increased while knockdown of OTUD6A decreased the CDC6 protein level in all three fractions (Fig. 1g and Supplementary Fig. 1m, n)."
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"Through a screening of DUBs, we found that OTUD6A interacts with CDC6 and increases the CDC6 protein level by promoting the stability of CDC6 through removing polyubiquitin chains, whose attachment was mediated by SCF-Cyclin F and APC/C-CDH1 (Fig. 8)."
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"Our results confirmed that the changes in the levels of CDC6 protein caused by OTUD6A were not results from changes in the cell cycle."
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"Here, we found that OTUD6A upregulated CDC6 protein level in several types of cancer cells."
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"Notably, knockout of OTUD6A resulted in less bladder tumorigenesis, lower malignancy and a lower CDC6 protein level upon BBN treatment (Fig. 5r-t), indicating that knockout of OTUD6A inhibited BCa tumorigenesis and CDC6 expression induced by chemical carcinogen."
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"The finding that OTUD6A increases the CDC6 protein level but does not affect the CDC6 mRNA level suggests that OTUD6A regulates CDC6 expression at the posttranscriptional level."
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"Overexpression of OTUD6A upregulated CDC6 protein level and prolonged the half-life of the CDC6 protein, independent of its phosphorylation status (Supplementary Fig. 3c-e)."
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"The reduced CDC6 protein level caused by OTUD6A knockdown was effectively restored by treatment with the proteasome inhibitor MG132 (Fig. 3d) but not with the lysosome inhibitor chloroquine (Supplementary Fig. 3f), indicating that OTUD6A likely maintains CDC6 protein stability through the proteasomal pathway."
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"In all cell lines detected, knockdown of OTUD6A decreased the protein level of CDC6, while the mRNA level was not affected (Fig. 5b and Supplementary Fig. 5a-e)."
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