Arrestin-3 scaffolding of the JNK3 cascade suggests a mechanism for signal amplification

Nicole A. Perry, Tamer S. Kaoud, Oscar O. Ortega, Ali I. Kaya, David J. Marcus, John M. Pleinis, Sandra Berndt, Qiuyan Chen, Xuanzhi Zhan, Kevin Dalby, Carlos F. Lopez, T. M. Iverson, Vsevolod V. Gurevich

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Scaffold proteins tether and orient components of a signaling cascade to facilitate signaling. Although much is known about how scaffolds colocalize signaling proteins, it is unclear whether scaffolds promote signal amplification. Here, we used arrestin-3, a scaffold of the ASK1-MKK4/7-JNK3 cascade, as a model to understand signal amplification by a scaffold protein. We found that arrestin-3 exhibited >15-fold higher affinity for inactive JNK3 than for active JNK3, and this change involved a shift in the binding site following JNK3 activation. We used systems biochemistry modeling and Bayesian inference to evaluate how the activation of upstream kinases contributed to JNK3 phosphorylation. Our combined experimental and computational approach suggested that the catalytic phosphorylation rate of JNK3 at Thr-221 by MKK7 is two orders of magnitude faster than the corresponding phosphorylation of Tyr-223 by MKK4 with or without arrestin-3. Finally, we showed that the release of activated JNK3 was critical for signal amplification. Collectively, our data suggest a “conveyor belt” mechanism for signal amplification by scaffold proteins. This mechanism informs on a long-standing mystery for how few upstream kinase molecules activate numerous downstream kinases to amplify signaling.

Original languageEnglish (US)
Pages (from-to)810-815
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume116
Issue number3
DOIs
StatePublished - Jan 15 2019

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Phosphotransferases
Phosphorylation
Proteins
Biochemistry
Binding Sites
beta-Arrestin 2

Keywords

  • Arrestin
  • Cell
  • Mitogen-activated protein kinase
  • Protein scaffold
  • Signal amplification
  • Signaling

ASJC Scopus subject areas

  • General

Cite this

Arrestin-3 scaffolding of the JNK3 cascade suggests a mechanism for signal amplification. / Perry, Nicole A.; Kaoud, Tamer S.; Ortega, Oscar O.; Kaya, Ali I.; Marcus, David J.; Pleinis, John M.; Berndt, Sandra; Chen, Qiuyan; Zhan, Xuanzhi; Dalby, Kevin; Lopez, Carlos F.; Iverson, T. M.; Gurevich, Vsevolod V.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 116, No. 3, 15.01.2019, p. 810-815.

Research output: Contribution to journalArticle

Perry, NA, Kaoud, TS, Ortega, OO, Kaya, AI, Marcus, DJ, Pleinis, JM, Berndt, S, Chen, Q, Zhan, X, Dalby, K, Lopez, CF, Iverson, TM & Gurevich, VV 2019, 'Arrestin-3 scaffolding of the JNK3 cascade suggests a mechanism for signal amplification', Proceedings of the National Academy of Sciences of the United States of America, vol. 116, no. 3, pp. 810-815. https://doi.org/10.1073/pnas.1819230116
Perry, Nicole A. ; Kaoud, Tamer S. ; Ortega, Oscar O. ; Kaya, Ali I. ; Marcus, David J. ; Pleinis, John M. ; Berndt, Sandra ; Chen, Qiuyan ; Zhan, Xuanzhi ; Dalby, Kevin ; Lopez, Carlos F. ; Iverson, T. M. ; Gurevich, Vsevolod V. / Arrestin-3 scaffolding of the JNK3 cascade suggests a mechanism for signal amplification. In: Proceedings of the National Academy of Sciences of the United States of America. 2019 ; Vol. 116, No. 3. pp. 810-815.
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