Tryptophan-mediated interactions between tristetraprolin and the CNOT9 subunit are required for CCR4-NOT deadenylase complex recruitment
The zinc-finger protein tristetraprolin (TTP) binds to AU-rich elements present in the 3' untranslated regions of transcripts that mainly encode proteins of the inflammatory response. TTP-bound mRNAs are targeted for destruction via recruitment of the eight-subunit deadenylase complex 'car...
| Main Authors: | , , , , , , , , , , , , , , , |
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| Format: | Journal article |
| Language: | English |
| Published: |
Elsevier
2017
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| _version_ | 1797085223608909824 |
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| author | Bulbrook, D Brazier, H Mahajan, P Kliszczak, M Fedorov, O Marchese, F Aubareda, A Chalk, R Picaud, S Strain-Damerell, C Filippakopoulos, P Gileadi, O Clark, A Yue, W Burgess-Brown, N Dean, J |
| author_facet | Bulbrook, D Brazier, H Mahajan, P Kliszczak, M Fedorov, O Marchese, F Aubareda, A Chalk, R Picaud, S Strain-Damerell, C Filippakopoulos, P Gileadi, O Clark, A Yue, W Burgess-Brown, N Dean, J |
| author_sort | Bulbrook, D |
| collection | OXFORD |
| description | The zinc-finger protein tristetraprolin (TTP) binds to AU-rich elements present in the 3' untranslated regions of transcripts that mainly encode proteins of the inflammatory response. TTP-bound mRNAs are targeted for destruction via recruitment of the eight-subunit deadenylase complex 'carbon catabolite repressor protein 4 (CCR4) -negative on TATA-less (NOT)' which catalyzes the removal of mRNA poly-(A) tails, the first obligatory step in mRNA decay. Here we show that a novel interaction between TTP and the CCR4-NOT subunit, CNOT9, is required for recruitment of the deadenylase complex. In addition to CNOT1, CNOT9 is now included in the identified CCR4-NOT subunits shown to interact with TTP. We find that both the N- and C-terminal domains of TTP are involved in an interaction with CNOT9. Through a combination of SPOT peptide array, site-directed mutagenesis and bio-layer interferometry, we identified several conserved tryptophan (Trp) residues in TTP that serve as major sites of interaction with two Trp-binding pockets of CNOT9, previously found to interact with another modulator GW182. We further demonstrate that these interactions are also required for recruitment of the CCR4-NOT complex and TTP-directed decay of an mRNA containing an AU-rich element in its 3'-untranslated region. Together the results reveal new molecular details for the TTP-CNOT interaction that shape an emerging mechanism whereby TTP targets inflammatory mRNAs for deadenylation and decay. |
| first_indexed | 2024-03-07T02:05:54Z |
| format | Journal article |
| id | oxford-uuid:9efc40d6-843f-447a-b601-5f0af21e223e |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T02:05:54Z |
| publishDate | 2017 |
| publisher | Elsevier |
| record_format | dspace |
| spelling | oxford-uuid:9efc40d6-843f-447a-b601-5f0af21e223e2022-03-27T00:54:07ZTryptophan-mediated interactions between tristetraprolin and the CNOT9 subunit are required for CCR4-NOT deadenylase complex recruitmentJournal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:9efc40d6-843f-447a-b601-5f0af21e223eEnglishSymplectic Elements at OxfordElsevier2017Bulbrook, DBrazier, HMahajan, PKliszczak, MFedorov, OMarchese, FAubareda, AChalk, RPicaud, SStrain-Damerell, CFilippakopoulos, PGileadi, OClark, AYue, WBurgess-Brown, NDean, JThe zinc-finger protein tristetraprolin (TTP) binds to AU-rich elements present in the 3' untranslated regions of transcripts that mainly encode proteins of the inflammatory response. TTP-bound mRNAs are targeted for destruction via recruitment of the eight-subunit deadenylase complex 'carbon catabolite repressor protein 4 (CCR4) -negative on TATA-less (NOT)' which catalyzes the removal of mRNA poly-(A) tails, the first obligatory step in mRNA decay. Here we show that a novel interaction between TTP and the CCR4-NOT subunit, CNOT9, is required for recruitment of the deadenylase complex. In addition to CNOT1, CNOT9 is now included in the identified CCR4-NOT subunits shown to interact with TTP. We find that both the N- and C-terminal domains of TTP are involved in an interaction with CNOT9. Through a combination of SPOT peptide array, site-directed mutagenesis and bio-layer interferometry, we identified several conserved tryptophan (Trp) residues in TTP that serve as major sites of interaction with two Trp-binding pockets of CNOT9, previously found to interact with another modulator GW182. We further demonstrate that these interactions are also required for recruitment of the CCR4-NOT complex and TTP-directed decay of an mRNA containing an AU-rich element in its 3'-untranslated region. Together the results reveal new molecular details for the TTP-CNOT interaction that shape an emerging mechanism whereby TTP targets inflammatory mRNAs for deadenylation and decay. |
| spellingShingle | Bulbrook, D Brazier, H Mahajan, P Kliszczak, M Fedorov, O Marchese, F Aubareda, A Chalk, R Picaud, S Strain-Damerell, C Filippakopoulos, P Gileadi, O Clark, A Yue, W Burgess-Brown, N Dean, J Tryptophan-mediated interactions between tristetraprolin and the CNOT9 subunit are required for CCR4-NOT deadenylase complex recruitment |
| title | Tryptophan-mediated interactions between tristetraprolin and the CNOT9 subunit are required for CCR4-NOT deadenylase complex recruitment |
| title_full | Tryptophan-mediated interactions between tristetraprolin and the CNOT9 subunit are required for CCR4-NOT deadenylase complex recruitment |
| title_fullStr | Tryptophan-mediated interactions between tristetraprolin and the CNOT9 subunit are required for CCR4-NOT deadenylase complex recruitment |
| title_full_unstemmed | Tryptophan-mediated interactions between tristetraprolin and the CNOT9 subunit are required for CCR4-NOT deadenylase complex recruitment |
| title_short | Tryptophan-mediated interactions between tristetraprolin and the CNOT9 subunit are required for CCR4-NOT deadenylase complex recruitment |
| title_sort | tryptophan mediated interactions between tristetraprolin and the cnot9 subunit are required for ccr4 not deadenylase complex recruitment |
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