IDEAS home Printed from https://ideas.repec.org/a/nat/natcom/v13y2022i1d10.1038_s41467-022-33762-3.html
   My bibliography  Save this article

Revealing β-TrCP activity dynamics in live cells with a genetically encoded biosensor

Author

Listed:
  • Debasish Paul

    (National Cancer Institute)

  • Stephen C. Kales

    (National Institutes of Health)

  • James A. Cornwell

    (National Cancer Institute)

  • Marwa M. Afifi

    (National Cancer Institute)

  • Ganesha Rai

    (National Institutes of Health)

  • Alexey Zakharov

    (National Institutes of Health)

  • Anton Simeonov

    (National Institutes of Health)

  • Steven D. Cappell

    (National Cancer Institute)

Abstract

The F-box protein beta-transducin repeat containing protein (β-TrCP) acts as a substrate adapter for the SCF E3 ubiquitin ligase complex, plays a crucial role in cell physiology, and is often deregulated in many types of cancers. Here, we develop a fluorescent biosensor to quantitatively measure β-TrCP activity in live, single cells in real-time. We find β-TrCP remains constitutively active throughout the cell cycle and functions to maintain discreet steady-state levels of its substrates. We find no correlation between expression levels of β-TrCP and β-TrCP activity, indicating post-transcriptional regulation. A high throughput screen of small-molecules using our reporter identifies receptor-tyrosine kinase signaling as a key axis for regulating β-TrCP activity by inhibiting binding between β-TrCP and the core SCF complex. Our study introduces a method to monitor β-TrCP activity in live cells and identifies a key signaling network that regulates β-TrCP activity throughout the cell cycle.

Suggested Citation

  • Debasish Paul & Stephen C. Kales & James A. Cornwell & Marwa M. Afifi & Ganesha Rai & Alexey Zakharov & Anton Simeonov & Steven D. Cappell, 2022. "Revealing β-TrCP activity dynamics in live cells with a genetically encoded biosensor," Nature Communications, Nature, vol. 13(1), pages 1-14, December.
    • Handle: RePEc:nat:natcom:v:13:y:2022:i:1:d:10.1038_s41467-022-33762-3
    DOI: 10.1038/s41467-022-33762-3
    as

    Download full text from publisher

    File URL: https://www.nature.com/articles/s41467-022-33762-3
    File Function: Abstract
    Download Restriction: no
    File URL: https://libkey.io/10.1038/s41467-022-33762-3?utm_source=ideas
    LibKey link: if access is restricted and if your library uses this service, LibKey will redirect you to where you can use your library subscription to access this item
    ---><---

    References listed on IDEAS

    as
    1. Teresa A. Soucy & Peter G. Smith & Michael A. Milhollen & Allison J. Berger & James M. Gavin & Sharmila Adhikari & James E. Brownell & Kristine E. Burke & David P. Cardin & Stephen Critchley & Courtne, 2009. "An inhibitor of NEDD8-activating enzyme as a new approach to treat cancer," Nature, Nature, vol. 458(7239), pages 732-736, April.
    2. Steven D. Cappell & Kevin G. Mark & Damien Garbett & Lindsey R. Pack & Michael Rape & Tobias Meyer, 2018. "EMI1 switches from being a substrate to an inhibitor of APC/CCDH1 to start the cell cycle," Nature, Nature, vol. 558(7709), pages 313-317, June.
    3. Thomas F. Westbrook & Guang Hu & Xiaolu L. Ang & Peter Mulligan & Natalya N. Pavlova & Anthony Liang & Yumei Leng & Rene Maehr & Yang Shi & J. Wade Harper & Stephen J. Elledge, 2008. "SCFβ-TRCP controls oncogenic transformation and neural differentiation through REST degradation," Nature, Nature, vol. 452(7185), pages 370-374, March.
    4. Hee Won Yang & Mingyu Chung & Takamasa Kudo & Tobias Meyer, 2017. "Competing memories of mitogen and p53 signalling control cell-cycle entry," Nature, Nature, vol. 549(7672), pages 404-408, September.
    5. Zhiwei Wang & Xiangpeng Dai & Jiateng Zhong & Hiroyuki Inuzuka & Lixin Wan & Xiaoning Li & Lixia Wang & Xiantao Ye & Liankun Sun & Daming Gao & Lee Zou & Wenyi Wei, 2015. "SCFβ-TRCP promotes cell growth by targeting PR-Set7/Set8 for degradation," Nature Communications, Nature, vol. 6(1), pages 1-11, December.
    Full references (including those not matched with items on IDEAS)

    Citations

    Citations are extracted by the CitEc Project, subscribe to its RSS feed for this item.
    as


    Cited by:

    1. Linda Lauinger & Anna Andronicos & Karin Flick & Clinton Yu & Geetha Durairaj & Lan Huang & Peter Kaiser, 2024. "Cadmium binding by the F-box domain induces p97-mediated SCF complex disassembly to activate stress response programs," Nature Communications, Nature, vol. 15(1), pages 1-13, December.

    Most related items

    These are the items that most often cite the same works as this one and are cited by the same works as this one.
    1. Yongchang Zhu & Xiping Zhang & Meng Gao & Yanchao Huang & Yuanqing Tan & Avital Parnas & Sizhong Wu & Delin Zhan & Sheera Adar & Jinchuan Hu, 2024. "Coordination of transcription-coupled repair and repair-independent release of lesion-stalled RNA polymerase II," Nature Communications, Nature, vol. 15(1), pages 1-15, December.
    2. María Arroyo & Florian D. Hastert & Andreas Zhadan & Florian Schelter & Susanne Zimbelmann & Cathia Rausch & Anne K. Ludwig & Thomas Carell & M. Cristina Cardoso, 2022. "Isoform-specific and ubiquitination dependent recruitment of Tet1 to replicating heterochromatin modulates methylcytosine oxidation," Nature Communications, Nature, vol. 13(1), pages 1-28, December.
    3. Sang Bae Lee & Luciano Garofano & Aram Ko & Fulvio D’Angelo & Brulinda Frangaj & Danika Sommer & Qiwen Gan & KyeongJin Kim & Timothy Cardozo & Antonio Iavarone & Anna Lasorella, 2022. "Regulated interaction of ID2 with the anaphase-promoting complex links progression through mitosis with reactivation of cell-type-specific transcription," Nature Communications, Nature, vol. 13(1), pages 1-14, December.
    4. Irineos Papakyriacou & Ginte Kutkaite & Marta Rúbies Bedós & Divya Nagarajan & Liam P. Alford & Michael P. Menden & Yumeng Mao, 2024. "Loss of NEDD8 in cancer cells causes vulnerability to immune checkpoint blockade in triple-negative breast cancer," Nature Communications, Nature, vol. 15(1), pages 1-19, December.
    5. Gang Xue & Jianing Xie & Matthias Hinterndorfer & Marko Cigler & Lara Dötsch & Hana Imrichova & Philipp Lampe & Xiufen Cheng & Soheila Rezaei Adariani & Georg E. Winter & Herbert Waldmann, 2023. "Discovery of a Drug-like, Natural Product-Inspired DCAF11 Ligand Chemotype," Nature Communications, Nature, vol. 14(1), pages 1-13, December.
    6. Shizhong Ke & Fabin Dang & Lin Wang & Jia-Yun Chen & Mandar T. Naik & Wenxue Li & Abhishek Thavamani & Nami Kim & Nandita M. Naik & Huaxiu Sui & Wei Tang & Chenxi Qiu & Kazuhiro Koikawa & Felipe Batal, 2024. "Reciprocal antagonism of PIN1-APC/CCDH1 governs mitotic protein stability and cell cycle entry," Nature Communications, Nature, vol. 15(1), pages 1-21, December.
    7. Daewon Lee & Eunju Yoon & Su Jin Ham & Kunwoo Lee & Hansaem Jang & Daihn Woo & Da Hyun Lee & Sehyeon Kim & Sekyu Choi & Jongkyeong Chung, 2024. "Diabetic sensory neuropathy and insulin resistance are induced by loss of UCHL1 in Drosophila," Nature Communications, Nature, vol. 15(1), pages 1-22, December.
    8. Andrea Riba & Attila Oravecz & Matej Durik & Sara Jiménez & Violaine Alunni & Marie Cerciat & Matthieu Jung & Céline Keime & William M. Keyes & Nacho Molina, 2022. "Cell cycle gene regulation dynamics revealed by RNA velocity and deep-learning," Nature Communications, Nature, vol. 13(1), pages 1-13, December.
    9. Nishtha Pandey & P K Vinod, 2018. "Mathematical modelling of reversible transition between quiescence and proliferation," PLOS ONE, Public Library of Science, vol. 13(6), pages 1-15, June.
    10. Jiajun Tan & Yingfeng Li & Xiang Li & Xiaoxiao Zhu & Liping Liu & Hua Huang & Jiahua Wei & Hailing Wang & Yong Tian & Zhigao Wang & Zhuqiang Zhang & Bing Zhu, 2024. "Pramel15 facilitates zygotic nuclear DNMT1 degradation and DNA demethylation," Nature Communications, Nature, vol. 15(1), pages 1-17, December.
    11. Mimi Zhang & Sungsoo Kim & Hee Won Yang, 2023. "Non-canonical pathway for Rb inactivation and external signaling coordinate cell-cycle entry without CDK4/6 activity," Nature Communications, Nature, vol. 14(1), pages 1-15, December.
    12. Di Wu & Haomin Li & Mingwei Liu & Jun Qin & Yi Sun, 2022. "The Ube2m-Rbx1 neddylation-Cullin-RING-Ligase proteins are essential for the maintenance of Regulatory T cell fitness," Nature Communications, Nature, vol. 13(1), pages 1-17, December.
    13. Yilun Sun & Simone A. Baechler & Xiaohu Zhang & Suresh Kumar & Valentina M. Factor & Yasuhiro Arakawa & Cindy H. Chau & Kanako Okamoto & Anup Parikh & Bob Walker & Yijun P. Su & Jiji Chen & Tabitha Ti, 2023. "Targeting neddylation sensitizes colorectal cancer to topoisomerase I inhibitors by inactivating the DCAF13-CRL4 ubiquitin ligase complex," Nature Communications, Nature, vol. 14(1), pages 1-20, December.
    14. Cheng Xu & Hongyi Zhou & Yulan Jin & Khushboo Sahay & Anna Robicsek & Yisong Liu & Kunzhe Dong & Jiliang Zhou & Amanda Barrett & Huabo Su & Weiqin Chen, 2022. "Hepatic neddylation deficiency triggers fatal liver injury via inducing NF-κB-inducing kinase in mice," Nature Communications, Nature, vol. 13(1), pages 1-18, December.
    15. Binod Kumar & Natania S. Field & Dale D. Kim & Asif A. Dar & Yanqun Chen & Aishwarya Suresh & Christopher F. Pastore & Li-Yin Hung & Nadia Porter & Keisuke Sawada & Palak Shah & Omar Elbulok & Emily K, 2022. "The ubiquitin ligase Cul5 regulates CD4+ T cell fate choice and allergic inflammation," Nature Communications, Nature, vol. 13(1), pages 1-14, December.
    16. Zack W Jones & Rachel Leander & Vito Quaranta & Leonard A Harris & Darren R Tyson, 2018. "A drift-diffusion checkpoint model predicts a highly variable and growth-factor-sensitive portion of the cell cycle G1 phase," PLOS ONE, Public Library of Science, vol. 13(2), pages 1-20, February.

    More about this item

    Statistics

    Access and download statistics

    Corrections

    All material on this site has been provided by the respective publishers and authors. You can help correct errors and omissions. When requesting a correction, please mention this item's handle: RePEc:nat:natcom:v:13:y:2022:i:1:d:10.1038_s41467-022-33762-3. See general information about how to correct material in RePEc.

    If you have authored this item and are not yet registered with RePEc, we encourage you to do it here. This allows to link your profile to this item. It also allows you to accept potential citations to this item that we are uncertain about.

    If CitEc recognized a bibliographic reference but did not link an item in RePEc to it, you can help with this form .

    If you know of missing items citing this one, you can help us creating those links by adding the relevant references in the same way as above, for each refering item. If you are a registered author of this item, you may also want to check the "citations" tab in your RePEc Author Service profile, as there may be some citations waiting for confirmation.

    For technical questions regarding this item, or to correct its authors, title, abstract, bibliographic or download information, contact: Sonal Shukla or Springer Nature Abstracting and Indexing (email available below). General contact details of provider: http://www.nature.com .

    Please note that corrections may take a couple of weeks to filter through the various RePEc services.

    IDEAS is a RePEc service. RePEc uses bibliographic data supplied by the respective publishers.