TY - JOUR
T1 - Targeted dephosphorylation of SMAD3 as an approach to impede TGFβ signaling
AU - Brewer, Abigail
AU - Zhao, Jin-Feng
AU - Fasimoye, Rotimi
AU - Shpiro, Natalia
AU - Macartney, Thomas J.
AU - Wood, Nicola
AU - Wightman, Melanie
AU - Alessi, Dario R.
AU - Sapkota, Gopal P.
N1 - Copyright:
© 2024 The Author(s). Published by Elsevier Inc.
PY - 2024/8/16
Y1 - 2024/8/16
N2 - TGFβ (transforming growth factor-β) signaling is involved in a myriad of cellular processes and its dysregulation has been implicated in many human diseases, including fibrosis and cancer. TGFβ transcriptional responses are controlled by tail phosphorylation of transcription factors SMAD2 and SMAD3 (mothers against decapentaplegic homolog 2/3). Therefore, targeted dephosphorylation of phospho-SMAD3 could provide an innovative mechanism to block some TGFβ-induced transcriptional responses, such as the transcription of SERPINE-1, which encodes plasminogen activator inhibitor 1 (PAI-1). Here, by developing and employing a bifunctional molecule, BDPIC (bromoTAG-dTAG proximity inducing chimera), we redirected multiple phosphatases, tagged with bromoTAG, to dephosphorylate phospho-SMAD3, tagged with dTAG. Using CRISPR/Cas9 technology, we generated homozygousdouble knock-in A549 bromoTAG/bromoTAGPPM1H/dTAG/dTAG SMAD3 cells, in which the BDPIC-induced proximity between bromoTAG-PPM1H and dTAG-SMAD3 led to a robust dephosphorylation of dTAG-SMAD3 and a significant decrease in SERPINE-1 transcription. Our work demonstrates targeted dephosphorylation of phospho-proteins as an exciting modality for rewiring cell signaling.
AB - TGFβ (transforming growth factor-β) signaling is involved in a myriad of cellular processes and its dysregulation has been implicated in many human diseases, including fibrosis and cancer. TGFβ transcriptional responses are controlled by tail phosphorylation of transcription factors SMAD2 and SMAD3 (mothers against decapentaplegic homolog 2/3). Therefore, targeted dephosphorylation of phospho-SMAD3 could provide an innovative mechanism to block some TGFβ-induced transcriptional responses, such as the transcription of SERPINE-1, which encodes plasminogen activator inhibitor 1 (PAI-1). Here, by developing and employing a bifunctional molecule, BDPIC (bromoTAG-dTAG proximity inducing chimera), we redirected multiple phosphatases, tagged with bromoTAG, to dephosphorylate phospho-SMAD3, tagged with dTAG. Using CRISPR/Cas9 technology, we generated homozygousdouble knock-in A549 bromoTAG/bromoTAGPPM1H/dTAG/dTAG SMAD3 cells, in which the BDPIC-induced proximity between bromoTAG-PPM1H and dTAG-SMAD3 led to a robust dephosphorylation of dTAG-SMAD3 and a significant decrease in SERPINE-1 transcription. Our work demonstrates targeted dephosphorylation of phospho-proteins as an exciting modality for rewiring cell signaling.
KW - Biochemistry
KW - Biochemistry methods
KW - Cell biology
UR - http://www.scopus.com/inward/record.url?scp=85198274891&partnerID=8YFLogxK
U2 - 10.1016/j.isci.2024.110423
DO - 10.1016/j.isci.2024.110423
M3 - Article
SN - 2589-0042
VL - 27
JO - iScience
JF - iScience
IS - 8
M1 - 110423
ER -
