Essential functions of the CNOT7/8 catalytic subunits of the CCR4-NOT complex in mRNA regulation and cell viability

• Dina, Mostafa

• Akinori, Takahashi

• Akiko, Yanagiya

• Tomokazu, Yamaguchi

• Takaya, Abe

• Taku, Kureha

• Keiji, Kuba

• Yumi, Kanegae

• Yasuhide, Furuta

• Tadashi, Yamamoto

• Toru, Suzuki

抄録

Shortening of mRNA poly(A) tails (deadenylation) to trigger their decay is mediated mainly by the CCR4-NOT deadenylase complex. While four catalytic subunits (CNOT6, 6L 7, and 8) have been identified in the mammalian CCR4-NOT complex, their individual biological roles are not fully understood. In this study, we addressed the contribution of CNOT7/8 to viability of primary mouse embryonic fibroblasts (MEFs). We found that MEFs lacking CNOT7/8 expression [Cnot7/8-double knockout (dKO) MEFs] undergo cell death, whereas MEFs lacking CNOT6/6L expression (Cnot6/6l-dKO MEFs) remain viable. Co-immunoprecipitation analyses showed that CNOT6/6L are also absent from the CCR4-NOT complex in Cnot7/8-dKO MEFs. In contrast, either CNOT7 or CNOT8 still interacts with other subunits in the CCR4-NOT complex in Cnot6/6l-dKO MEFs. Exogenous expression of a CNOT7 mutant lacking catalytic activity in Cnot7/8-dKO MEFs cannot recover cell viability, even though CNOT6/6L exists to some extent in the CCR4-NOT complex, confirming that CNOT7/8 is essential for viability. Bulk poly(A) tail analysis revealed that mRNAs with longer poly(A) tails are more numerous in Cnot7/8-dKO MEFs than in Cnot6/6l-dKO MEFs. Consistent with elongated poly(A) tails, more mRNAs are upregulated and stabilized in Cnot7/8-dKO MEFs than in Cnot6/6l-dKO MEFs. Importantly, Cnot6/6l-dKO mice are viable and grow normally to adulthood. Taken together, the CNOT7/8 catalytic subunits are essential for deadenylation, which is necessary to maintain cell viability, whereas CNOT6/6L are not.

source:http://creativecommons.org/licenses/by-nc-nd/4.0/

source:https://www.tandfonline.com/doi/full/10.1080/15476286.2019.1709747?scroll=top&needAccess=true

収録刊行物

• RNA Biology

  RNA Biology 17 (3), 403-416, 2020-01-10

  Informa UK Limited, trading as Taylor & Francis Group.

キーワード

• Cell Biology
• Molecular Biology

詳細情報

• CRID: 1050566774891551232
• NII論文ID: 120006840050
• ISSN: 15476286
• ISSN: 15558584
• DOI: 10.1080/15476286.2019.1709747
• Web Site: http://id.nii.ac.jp/1394/00001097/
• Web Site: https://www.tandfonline.com/doi/pdf/10.1080/15476286.2019.1709747
• 本文言語コード: en
• 資料種別: journal article
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