TY - JOUR
T1 - Extracellular matrix stiffness regulates degradation of MST2 via SCF βTrCP
AU - Fiore, Ana Paula Zen Petisco
AU - Rodrigues, Ana Maria
AU - Ribeiro-Filho, Helder Veras
AU - Manucci, Antonio Carlos
AU - de Freitas Ribeiro, Pedro
AU - Botelho, Mayara Carolinne Silva
AU - Vogel, Christine
AU - Lopes-de-Oliveira, Paulo Sergio
AU - Pagano, Michele
AU - Bruni-Cardoso, Alexandre
N1 - Funding Information:
This work was supported by Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Young Investigator Award (14/10492-0), by a FAPESP grant (2019/26767-2), by the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq-Universal 444597/2014-0) and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES, Finance Code 001). MP is an investigator with the Howard Hughes Medical Institute and his lab is partially funded by grants from the National Institute of Health (R01-CA76584 and R35-GM136250). APZPF is a FAPESP Postdoctoral fellowship awardee (14/25832-1 and 17/18641-3). AMRS is funded by CAPES PhD-scholarship (PROEX), PFR is a FAPESP PhD-scholarship recipient (17/18067-5), MCSB is a FAPESP PhD-scholarship recipient (2017/25437-3) and ACM is a CNPq PhD-scholarship recipient (14668/2019-9). PLSO and HVRF are funded by a FAPESP grant (2018/00629-0). CV acknowledges funding by National Institute of Health (R35-GM127089). The authors would like to thank Dr. Gergely Róna (Department of Biochemistry and Molecular Pharmacology, New York University School of Medicine), Dr. Rebeka Tomasin (DBQ-IQUSP), Dr. Deborah Schechtman (DBQ-IQUSP), Dr. Marcelo Damário Gomes (Faculdade de Medicina de Ribeirão Preto – Universidade de São Paulo) and Dr. Hernandes F. Carvalho (UNICAMP) for all the fruitful discussions and valuable advices during the progress of this project. The authors would also like to thank Jeffrey Estrada, Celia Ludio Braga, Maria Luiza Baldini and Izaura Nobuko Toma for technical assistance.
Funding Information:
This work was supported by Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Young Investigator Award ( 14/10492-0 ), by a FAPESP grant ( 2019/26767-2 ), by the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq-Universal 444597/2014-0 ) and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES, Finance Code 001). MP is an investigator with the Howard Hughes Medical Institute and his lab is partially funded by grants from the National Institute of Health ( R01-CA76584 and R35-GM136250 ). APZPF is a FAPESP Postdoctoral fellowship awardee (14/25832-1 and 17/18641-3). AMRS is funded by CAPES PhD-scholarship (PROEX), PFR is a FAPESP PhD-scholarship recipient (17/18067-5), MCSB is a FAPESP PhD-scholarship recipient (2017/25437-3) and ACM is a CNPq PhD-scholarship recipient (14668/2019-9). PLSO and HVRF are funded by a FAPESP grant (2018/00629-0). CV acknowledges funding by National Institute of Health ( R35-GM127089 ). The authors would like to thank Dr. Gergely Róna ( Department of Biochemistry and Molecular Pharmacology, New York University School of Medicine ), Dr. Rebeka Tomasin (DBQ-IQUSP), Dr. Deborah Schechtman (DBQ-IQUSP), Dr. Marcelo Damário Gomes ( Faculdade de Medicina de Ribeirão Preto – Universidade de São Paulo ) and Dr. Hernandes F. Carvalho (UNICAMP) for all the fruitful discussions and valuable advices during the progress of this project. The authors would also like to thank Jeffrey Estrada, Celia Ludio Braga, Maria Luiza Baldini and Izaura Nobuko Toma for technical assistance.
Publisher Copyright:
© 2022 Elsevier B.V.
PY - 2022/12
Y1 - 2022/12
N2 - The Hippo pathway plays central roles in relaying mechanical signals during development and tumorigenesis, but how the proteostasis of the Hippo kinase MST2 is regulated remains unknown. Here, we found that chemical inhibition of proteasomal proteolysis resulted in increased levels of MST2 in human breast epithelial cells. MST2 binds SCFβTrCP E3 ubiquitin ligase and silencing βTrCP resulted in MST2 accumulation. Site-directed mutagenesis combined with computational molecular dynamics studies revealed that βTrCP binds MST2 via a non-canonical degradation motif. Additionally, stiffer extracellular matrix, as well as hyperactivation of integrins resulted in enhanced MST2 degradation mediated by integrin-linked kinase (ILK) and actomyosin stress fibers. Our study uncovers the underlying biochemical mechanisms controlling MST2 degradation and underscores how alterations in the microenvironment rigidity regulate the proteostasis of a central Hippo pathway component.
AB - The Hippo pathway plays central roles in relaying mechanical signals during development and tumorigenesis, but how the proteostasis of the Hippo kinase MST2 is regulated remains unknown. Here, we found that chemical inhibition of proteasomal proteolysis resulted in increased levels of MST2 in human breast epithelial cells. MST2 binds SCFβTrCP E3 ubiquitin ligase and silencing βTrCP resulted in MST2 accumulation. Site-directed mutagenesis combined with computational molecular dynamics studies revealed that βTrCP binds MST2 via a non-canonical degradation motif. Additionally, stiffer extracellular matrix, as well as hyperactivation of integrins resulted in enhanced MST2 degradation mediated by integrin-linked kinase (ILK) and actomyosin stress fibers. Our study uncovers the underlying biochemical mechanisms controlling MST2 degradation and underscores how alterations in the microenvironment rigidity regulate the proteostasis of a central Hippo pathway component.
KW - Breast cells
KW - Extracellular matrix stiffness
KW - Hippo
KW - MST2
KW - SCF βTrCP
KW - Ubiquitinproteasome system
UR - http://www.scopus.com/inward/record.url?scp=85137013831&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85137013831&partnerID=8YFLogxK
U2 - 10.1016/j.bbagen.2022.130238
DO - 10.1016/j.bbagen.2022.130238
M3 - Article
C2 - 36044955
AN - SCOPUS:85137013831
SN - 0304-4165
VL - 1866
JO - Biochimica et Biophysica Acta - General Subjects
JF - Biochimica et Biophysica Acta - General Subjects
IS - 12
M1 - 130238
ER -