IndraLab

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OTUD5 binds SUPT16H. 14 / 19
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"To study the cellular consequence of specifically blocking the interaction between OTUD5 and FACT (OTUD5 overexpression or complete knockout causes aberration in cell cycle and apoptosis), we generated a HeLa knock-in (KI) cell line in which the D537A mutation is introduced into the genomic loci of OTUD5 (OTUD5 ) using the CRISPR-Cas9 method (Supplementary Figure S2; sequencing information of 4 clones)."
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"Thus, blocking the interaction between FACT and OTUD5 enhances FACT nucleosome remodeling function.The OTUD5 cells display a noticeable increase in micronuclei and cytoplasmic bridges (Figure 3E), which can form as a consequence of replication stress."
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"In addition, OTUD5 can interact with the SPT16 protein in the FACT complex and antagonize the deposition of histone H2A upon DSB damage by acting as a scaffold [ xref , xref ] ( xref D)."
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"OTUD5 interacts with SPT16 and regulates the SPT16-dependent Pol II elongation at damaged chromatin."
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"To test whether OTUD5 also interacts with SPT16 in an unbiased method, we performed FLAG-OTUD5 IP and GST-OTUD5 pulldown experiments from HeLa S3 cells and subjected the eluate to mass spec analysis (Figure xref )."
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"Further, the interaction between OTUD5 and SPT16 can be detected in situ by the PLA (Figure xref )."
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"These results unequivocally suggest that both the UBR5–OTUD5 interaction and OTUD5–SPT16 interaction are necessary for transcriptional repression at the damaged chromatin."
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"Independently of UBR5, OTUD5 also interacts with SPT16, through the UIM domain at the C-terminal tail."
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"These results confirm that the interaction between OTUD5 and FACT is important for the transcriptional regulation, and provide a new possible link between the FACT-dependent transcriptional process and tumor suppression."
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"Further investigation into the nature of the interaction between OTUD5 and FACT will provide more insight into the regulation."
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"We speculate that the OTUD5–SPT16 interaction, which occurs through the UIM domain, may pose an additional role in the repression of cytokine production described in the earlier studies."
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"Disruption of the OTUD5-SPT16 interaction was also confirmed using PLA wherein the PLA signals were undetectable in the nucleus of KI cells (Figure xref )."
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"Independently from its interaction with UBR5, OTUD5 also interacts with SPT16, another component of the FACT complex."
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"In addition, OTUD5 interacts with the FACT component SPT16 and antagonizes histone H2A deposition at DSBs [ xref ]."
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