Proteomics

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The exon junction complex component CASC3 is a major effector of nonsense-mediated mRNA decay in human cells

ABSTRACT: In metazoans, the exon junction complex (EJC) is a central component of spliced messenger ribonucleoprotein particles (mRNPs). CASC3 has been reported to be a core component of the EJC and to be crucial for assembly, the splicing regulating function of the EJC and nonsense-mediated mRNA decay (NMD). However, recent evidence suggests that CASC3 functions differently from other EJC core components. To elucidate the cellular role of CASC3, we have established human cell lines in which CASC3 was inactivated by means of CRISPR-Cas9 genome editing. We show that in these cells CASC3 is dispensable for the splicing regulatory role of the EJC. However, we find that CASC3 depletion results in the upregulation of many known and novel NMD substrates, suggesting that CASC3 is required for the efficient execution of EJC-dependent NMD. Taken together, our results challenge the model of CASC3 as an assembly factor and core component of the EJC. Our data rather show that CASC3 is involved in the degradation of NMD substrates and therefore uncover the primary molecular function of CASC3 in human cells.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture, Kidney Cell

SUBMITTER: Janica Wiederstein  

LAB HEAD: Niels H. Gehring

PROVIDER: PXD019161 | Pride | 2021-09-09

REPOSITORIES: Pride

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20200414_Q4_SK_ColID_968_Jenny_Gehring_IP_FLAG-CASC3_01.raw Raw
20200414_Q4_SK_ColID_968_Jenny_Gehring_IP_FLAG-CASC3_02.raw Raw
20200414_Q4_SK_ColID_968_Jenny_Gehring_IP_FLAG-CASC3_03.raw Raw
20200414_Q4_SK_ColID_968_Jenny_Gehring_IP_FLAG-CASC3_04.raw Raw
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The exon junction complex (EJC) is an essential constituent and regulator of spliced messenger ribonucleoprotein particles (mRNPs) in metazoans. As a core component of the EJC, CASC3 was described to be pivotal for EJC-dependent nuclear and cytoplasmic processes. However, recent evidence suggests that CASC3 functions differently from other EJC core proteins. Here, we have established human CASC3 knockout cell lines to elucidate the cellular role of CASC3. In the knockout cells, overall EJC compo  ...[more]

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