Tryptophan-Mediated Interactions between Tristetraprolin and the CNOT9 Subunit Are Required for CCR4-NOT Deadenylase Complex Recruitment

Research output: Contribution to journalArticlepeer-review

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Tryptophan-Mediated Interactions between Tristetraprolin and the CNOT9 Subunit Are Required for CCR4-NOT Deadenylase Complex Recruitment. / Bulbrook, D.; Brazier, H.; Mahajan, P.; Kliszczak, M.; Fedorov, O.; Marchese, F. P.; Aubareda, A.; Chalk, R.; Picaud, S.; Strain-Damerell, C.; Filippakopoulos, P.; Gileadi, O.; Clark, A. R.; Yue, W.w.; Burgess-Brown, N. A.; Dean, J. L. E.

In: Journal of Molecular Biology, 29.12.2017.

Research output: Contribution to journalArticlepeer-review

Harvard

Bulbrook, D, Brazier, H, Mahajan, P, Kliszczak, M, Fedorov, O, Marchese, FP, Aubareda, A, Chalk, R, Picaud, S, Strain-Damerell, C, Filippakopoulos, P, Gileadi, O, Clark, AR, Yue, WW, Burgess-Brown, NA & Dean, JLE 2017, 'Tryptophan-Mediated Interactions between Tristetraprolin and the CNOT9 Subunit Are Required for CCR4-NOT Deadenylase Complex Recruitment', Journal of Molecular Biology. https://doi.org/10.1016/j.jmb.2017.12.018

APA

Bulbrook, D., Brazier, H., Mahajan, P., Kliszczak, M., Fedorov, O., Marchese, F. P., Aubareda, A., Chalk, R., Picaud, S., Strain-Damerell, C., Filippakopoulos, P., Gileadi, O., Clark, A. R., Yue, W. W., Burgess-Brown, N. A., & Dean, J. L. E. (2017). Tryptophan-Mediated Interactions between Tristetraprolin and the CNOT9 Subunit Are Required for CCR4-NOT Deadenylase Complex Recruitment. Journal of Molecular Biology. https://doi.org/10.1016/j.jmb.2017.12.018

Vancouver

Author

Bulbrook, D. ; Brazier, H. ; Mahajan, P. ; Kliszczak, M. ; Fedorov, O. ; Marchese, F. P. ; Aubareda, A. ; Chalk, R. ; Picaud, S. ; Strain-Damerell, C. ; Filippakopoulos, P. ; Gileadi, O. ; Clark, A. R. ; Yue, W.w. ; Burgess-Brown, N. A. ; Dean, J. L. E. / Tryptophan-Mediated Interactions between Tristetraprolin and the CNOT9 Subunit Are Required for CCR4-NOT Deadenylase Complex Recruitment. In: Journal of Molecular Biology. 2017.

Bibtex

@article{56aabc2dec6446f4aa4fb2a4acc2032f,
title = "Tryptophan-Mediated Interactions between Tristetraprolin and the CNOT9 Subunit Are Required for CCR4-NOT Deadenylase Complex Recruitment",
abstract = "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 residues in TTP that serve as major sites of interaction with two tryptophan-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.",
keywords = "deadenylation , inflammatory , mRNA , AU-rich elements , post-translational controls",
author = "D. Bulbrook and H. Brazier and P. Mahajan and M. Kliszczak and O. Fedorov and Marchese, {F. P.} and A. Aubareda and R. Chalk and S. Picaud and C. Strain-Damerell and P. Filippakopoulos and O. Gileadi and Clark, {A. R.} and W.w. Yue and Burgess-Brown, {N. A.} and Dean, {J. L. E.}",
year = "2017",
month = dec,
day = "29",
doi = "10.1016/j.jmb.2017.12.018",
language = "English",
journal = "Journal of Molecular Biology",
issn = "0022-2836",
publisher = "Elsevier",

}

RIS

TY - JOUR

T1 - Tryptophan-Mediated Interactions between Tristetraprolin and the CNOT9 Subunit Are Required for CCR4-NOT Deadenylase Complex Recruitment

AU - Bulbrook, D.

AU - Brazier, H.

AU - Mahajan, P.

AU - Kliszczak, M.

AU - Fedorov, O.

AU - Marchese, F. P.

AU - Aubareda, A.

AU - Chalk, R.

AU - Picaud, S.

AU - Strain-Damerell, C.

AU - Filippakopoulos, P.

AU - Gileadi, O.

AU - Clark, A. R.

AU - Yue, W.w.

AU - Burgess-Brown, N. A.

AU - Dean, J. L. E.

PY - 2017/12/29

Y1 - 2017/12/29

N2 - 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 residues in TTP that serve as major sites of interaction with two tryptophan-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.

AB - 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 residues in TTP that serve as major sites of interaction with two tryptophan-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.

KW - deadenylation

KW - inflammatory

KW - mRNA

KW - AU-rich elements

KW - post-translational controls

U2 - 10.1016/j.jmb.2017.12.018

DO - 10.1016/j.jmb.2017.12.018

M3 - Article

JO - Journal of Molecular Biology

JF - Journal of Molecular Biology

SN - 0022-2836

ER -