Receptor tyrosine kinases activate heterotrimeric G proteins via phosphorylation within the interdomain cleft of Gαi

Nicholas A Kalogriopoulos; Inmaculada Lopez-Sanchez; Changsheng Lin; Tony Ngo; Krishna K Midde; Suchismita Roy; Nicolas Aznar; Fiona Murray; Mikel Garcia-Marcos; Irina Kufareva; Majid Ghassemian; Pradipta Ghosh

doi:10.1073/pnas.2004699117

Receptor tyrosine kinases activate heterotrimeric G proteins via phosphorylation within the interdomain cleft of Gαi

Nicholas A Kalogriopoulos, Inmaculada Lopez-Sanchez, Changsheng Lin, Tony Ngo, Krishna K Midde, Suchismita Roy, Nicolas Aznar, Fiona Murray, Mikel Garcia-Marcos, Irina Kufareva, Majid Ghassemian, Pradipta Ghosh^* (Corresponding Author)

^*Corresponding author for this work

Medical Sciences

Research output: Contribution to journal › Article › peer-review

13 Citations (Scopus)

Abstract

The molecular mechanisms by which receptor tyrosine kinases (RTKs) and heterotrimeric G proteins, two major signaling hubs in eukaryotes, independently relay signals across the plasma membrane have been extensively characterized. How these hubs cross-talk has been a long-standing question, but answers remain elusive. Using linear ion-trap mass spectrometry in combination with biochemical, cellular, and computational approaches, we unravel a mechanism of activation of heterotrimeric G proteins by RTKs and chart the key steps that mediate such activation. Upon growth factor stimulation, the guanine-nucleotide exchange modulator dissociates Gαi•βγ trimers, scaffolds monomeric Gαi with RTKs, and facilitates the phosphorylation on two tyrosines located within the interdomain cleft of Gαi. Phosphorylation triggers the activation of Gαi and inhibits second messengers (cAMP). Tumor-associated mutants reveal how constitutive activation of this pathway impacts cell's decision to "go" vs. "grow." These insights define a tyrosine-based G protein signaling paradigm and reveal its importance in eukaryotes.

Original language	English
Pages (from-to)	28763-28774
Number of pages	12
Journal	Proceedings of the National Academy of Sciences of the United States of America
Volume	117
Issue number	46
Early online date	17 Nov 2020
DOIs	https://doi.org/10.1073/pnas.2004699117
Publication status	Published - 17 Nov 2020

Bibliographical note

Acknowledgments

We thank Bridgett Simmons (AB SCIEX) for technical assistance with mass spectrometry experiments, and Yelena Pavlova and Nina Sun for technical assistance with cloning and mutagenesis of constructs. This paper was supported by the NIH Grants CA238042, AI141630, CA100768, and CA160911 (to P.G.). N.A.K. was supported by an NIH predoctoral fellowship (F31 CA206426), and T32 training Grants T32CA067754 and T32DK007202. M.G.-M. was supported by the NIH (GM136132 and GM130120). I.L.-S. was supported by a fellowship from the American Heart Association (AHA 14POST20050025). I.K. was supported by the NIH (AI118985 and R01 GM117424). T.N. is supported by National Health and Meducak Research Council C. J. Martin Early Career Fellowship 1145746.

Keywords

heterotrimeric G proteins
growth factor receptor tyrosine kinases
EGFR
tyrosine
phosphorylation
transactivation

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Fiona Murray
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Kalogriopoulos, N. A., Lopez-Sanchez, I., Lin, C., Ngo, T., Midde, K. K., Roy, S., Aznar, N., Murray, F., Garcia-Marcos, M., Kufareva, I., Ghassemian, M., & Ghosh, P. (2020). Receptor tyrosine kinases activate heterotrimeric G proteins via phosphorylation within the interdomain cleft of Gαi. Proceedings of the National Academy of Sciences of the United States of America, 117(46), 28763-28774. https://doi.org/10.1073/pnas.2004699117

Receptor tyrosine kinases activate heterotrimeric G proteins via phosphorylation within the interdomain cleft of Gαi. / Kalogriopoulos, Nicholas A; Lopez-Sanchez, Inmaculada; Lin, Changsheng et al.
In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 117, No. 46, 17.11.2020, p. 28763-28774.

Research output: Contribution to journal › Article › peer-review

Kalogriopoulos, NA, Lopez-Sanchez, I, Lin, C, Ngo, T, Midde, KK, Roy, S, Aznar, N, Murray, F, Garcia-Marcos, M, Kufareva, I, Ghassemian, M & Ghosh, P 2020, 'Receptor tyrosine kinases activate heterotrimeric G proteins via phosphorylation within the interdomain cleft of Gαi', Proceedings of the National Academy of Sciences of the United States of America, vol. 117, no. 46, pp. 28763-28774. https://doi.org/10.1073/pnas.2004699117

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note = "Acknowledgments We thank Bridgett Simmons (AB SCIEX) for technical assistance with mass spectrometry experiments, and Yelena Pavlova and Nina Sun for technical assistance with cloning and mutagenesis of constructs. This paper was supported by the NIH Grants CA238042, AI141630, CA100768, and CA160911 (to P.G.). N.A.K. was supported by an NIH predoctoral fellowship (F31 CA206426), and T32 training Grants T32CA067754 and T32DK007202. M.G.-M. was supported by the NIH (GM136132 and GM130120). I.L.-S. was supported by a fellowship from the American Heart Association (AHA 14POST20050025). I.K. was supported by the NIH (AI118985 and R01 GM117424). T.N. is supported by National Health and Meducak Research Council C. J. Martin Early Career Fellowship 1145746.",

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AU - Kalogriopoulos, Nicholas A

AU - Lopez-Sanchez, Inmaculada

AU - Lin, Changsheng

AU - Ngo, Tony

AU - Midde, Krishna K

AU - Roy, Suchismita

AU - Aznar, Nicolas

AU - Murray, Fiona

AU - Garcia-Marcos, Mikel

AU - Kufareva, Irina

AU - Ghassemian, Majid

AU - Ghosh, Pradipta

N1 - Acknowledgments We thank Bridgett Simmons (AB SCIEX) for technical assistance with mass spectrometry experiments, and Yelena Pavlova and Nina Sun for technical assistance with cloning and mutagenesis of constructs. This paper was supported by the NIH Grants CA238042, AI141630, CA100768, and CA160911 (to P.G.). N.A.K. was supported by an NIH predoctoral fellowship (F31 CA206426), and T32 training Grants T32CA067754 and T32DK007202. M.G.-M. was supported by the NIH (GM136132 and GM130120). I.L.-S. was supported by a fellowship from the American Heart Association (AHA 14POST20050025). I.K. was supported by the NIH (AI118985 and R01 GM117424). T.N. is supported by National Health and Meducak Research Council C. J. Martin Early Career Fellowship 1145746.

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N2 - The molecular mechanisms by which receptor tyrosine kinases (RTKs) and heterotrimeric G proteins, two major signaling hubs in eukaryotes, independently relay signals across the plasma membrane have been extensively characterized. How these hubs cross-talk has been a long-standing question, but answers remain elusive. Using linear ion-trap mass spectrometry in combination with biochemical, cellular, and computational approaches, we unravel a mechanism of activation of heterotrimeric G proteins by RTKs and chart the key steps that mediate such activation. Upon growth factor stimulation, the guanine-nucleotide exchange modulator dissociates Gαi•βγ trimers, scaffolds monomeric Gαi with RTKs, and facilitates the phosphorylation on two tyrosines located within the interdomain cleft of Gαi. Phosphorylation triggers the activation of Gαi and inhibits second messengers (cAMP). Tumor-associated mutants reveal how constitutive activation of this pathway impacts cell's decision to "go" vs. "grow." These insights define a tyrosine-based G protein signaling paradigm and reveal its importance in eukaryotes.

AB - The molecular mechanisms by which receptor tyrosine kinases (RTKs) and heterotrimeric G proteins, two major signaling hubs in eukaryotes, independently relay signals across the plasma membrane have been extensively characterized. How these hubs cross-talk has been a long-standing question, but answers remain elusive. Using linear ion-trap mass spectrometry in combination with biochemical, cellular, and computational approaches, we unravel a mechanism of activation of heterotrimeric G proteins by RTKs and chart the key steps that mediate such activation. Upon growth factor stimulation, the guanine-nucleotide exchange modulator dissociates Gαi•βγ trimers, scaffolds monomeric Gαi with RTKs, and facilitates the phosphorylation on two tyrosines located within the interdomain cleft of Gαi. Phosphorylation triggers the activation of Gαi and inhibits second messengers (cAMP). Tumor-associated mutants reveal how constitutive activation of this pathway impacts cell's decision to "go" vs. "grow." These insights define a tyrosine-based G protein signaling paradigm and reveal its importance in eukaryotes.

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KW - EGFR

KW - tyrosine

KW - phosphorylation

KW - transactivation

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DO - 10.1073/pnas.2004699117

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JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

IS - 46

ER -

Receptor tyrosine kinases activate heterotrimeric G proteins via phosphorylation within the interdomain cleft of Gαi

Abstract

Bibliographical note

Keywords

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