Structure of a PKA RIα Recurrent Acrodysostosis Mutant Explains Defective cAMP-Dependent Activation
Abstract
Most disease related mutations that impair PKA signaling are present within the regulatory PKA RI alpha-subunit (RIα). Although mutations in the PRKAR1A gene are linked to Carney complex disease (CNC) and more recently to acrodysostosis-1 (ACRDYS1), the two diseases show contrasting phenotypes. While CNC mutations cause increased PKA activity, ACRDYS1 mutations result in decreased PKA activity and cAMP resistant holoenzymes. Mapping the ACRDYS1 disease mutations reveals their localization to the second of two tandem cAMP binding domains (CNB-B) and here we characterize a recurrent deletion mutant where the last 14 residues are missing. The crystal structure of a monomeric form of this mutant (RIα92-365) bound to the C subunit reveals the dysfunctional regions of the RIα-subunit. Beyond the missing residues, the entire capping motif is disordered (residues 357-379) and explains the disrupted cAMP binding. Moreover, the effects of the mutation extend far beyond the CNB-B domain and include the active site and N-lobe of the C-subunit, which is in a partially open conformation with the C-tail disordered. A key residue that contributes to this crosstalk, D267, is altered in our structure and we confirmed its functional importance by mutagenesis. In particular, the D267 interaction with Arg241, a residue shown earliermore »
- Authors:
-
- Univ. of California at San Diego, La Jolla, CA (United States)
- Univ. of Utah, Salt Lake City, UT (United States)
- Univ. of Innsbruck, Innsbruck (Austria)
- Publication Date:
- Research Org.:
- Univ. of Utah, Salt Lake City, UT (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC), Basic Energy Sciences (BES)
- OSTI Identifier:
- 1422048
- Alternate Identifier(s):
- OSTI ID: 1410896
- Grant/Contract Number:
- FG02-05ER64026; AC02-05CH11231
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Journal of Molecular Biology
- Additional Journal Information:
- Journal Volume: 428; Journal Issue: PB; Journal ID: ISSN 0022-2836
- Publisher:
- Elsevier
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; PKA signaling; RIα subunit; disease mutations; crystal structure
Citation Formats
Bruystens, Jessica G. H., Wu, Jian, Fortezzo, Audrey, Del Rio, Jason, Nielsen, Cole, Blumenthal, Donald K., Rock, Ruth, Stefan, Eduard, and Taylor, Susan S. Structure of a PKA RIα Recurrent Acrodysostosis Mutant Explains Defective cAMP-Dependent Activation. United States: N. p., 2016.
Web. doi:10.1016/j.jmb.2016.10.033.
Bruystens, Jessica G. H., Wu, Jian, Fortezzo, Audrey, Del Rio, Jason, Nielsen, Cole, Blumenthal, Donald K., Rock, Ruth, Stefan, Eduard, & Taylor, Susan S. Structure of a PKA RIα Recurrent Acrodysostosis Mutant Explains Defective cAMP-Dependent Activation. United States. https://doi.org/10.1016/j.jmb.2016.10.033
Bruystens, Jessica G. H., Wu, Jian, Fortezzo, Audrey, Del Rio, Jason, Nielsen, Cole, Blumenthal, Donald K., Rock, Ruth, Stefan, Eduard, and Taylor, Susan S. Sat .
"Structure of a PKA RIα Recurrent Acrodysostosis Mutant Explains Defective cAMP-Dependent Activation". United States. https://doi.org/10.1016/j.jmb.2016.10.033. https://www.osti.gov/servlets/purl/1422048.
@article{osti_1422048,
title = {Structure of a PKA RIα Recurrent Acrodysostosis Mutant Explains Defective cAMP-Dependent Activation},
author = {Bruystens, Jessica G. H. and Wu, Jian and Fortezzo, Audrey and Del Rio, Jason and Nielsen, Cole and Blumenthal, Donald K. and Rock, Ruth and Stefan, Eduard and Taylor, Susan S.},
abstractNote = {Most disease related mutations that impair PKA signaling are present within the regulatory PKA RI alpha-subunit (RIα). Although mutations in the PRKAR1A gene are linked to Carney complex disease (CNC) and more recently to acrodysostosis-1 (ACRDYS1), the two diseases show contrasting phenotypes. While CNC mutations cause increased PKA activity, ACRDYS1 mutations result in decreased PKA activity and cAMP resistant holoenzymes. Mapping the ACRDYS1 disease mutations reveals their localization to the second of two tandem cAMP binding domains (CNB-B) and here we characterize a recurrent deletion mutant where the last 14 residues are missing. The crystal structure of a monomeric form of this mutant (RIα92-365) bound to the C subunit reveals the dysfunctional regions of the RIα-subunit. Beyond the missing residues, the entire capping motif is disordered (residues 357-379) and explains the disrupted cAMP binding. Moreover, the effects of the mutation extend far beyond the CNB-B domain and include the active site and N-lobe of the C-subunit, which is in a partially open conformation with the C-tail disordered. A key residue that contributes to this crosstalk, D267, is altered in our structure and we confirmed its functional importance by mutagenesis. In particular, the D267 interaction with Arg241, a residue shown earlier to be important for allosteric regulation, is disrupted thereby strengthening the interaction of D267 with the C-subunit residue Arg194 at the R:C interface. Here, we see here how the switch between active (cAMP-bound) and inactive (holoenzyme) conformations is perturbed and how the dynamically controlled crosstalk between the helical domains of the two CNB-domains is necessary for functional regulation of PKA activity.},
doi = {10.1016/j.jmb.2016.10.033},
journal = {Journal of Molecular Biology},
number = PB,
volume = 428,
place = {United States},
year = {Sat Nov 05 00:00:00 EDT 2016},
month = {Sat Nov 05 00:00:00 EDT 2016}
}
Web of Science
Works referenced in this record:
PKA-I Holoenzyme Structure Reveals a Mechanism for cAMP-Dependent Activation
journal, September 2007
- Kim, Choel; Cheng, Cecilia Y.; Saldanha, S. Adrian
- Cell, Vol. 130, Issue 6
Crystal Structure of a Complex Between the Catalytic and Regulatory (RI ) Subunits of PKA
journal, February 2005
- Kim, C.
- Science, Vol. 307, Issue 5710
The tissue-specific extinguisher locus TSE1 encodes a regulatory subunit of cAMP-Dependent protein kinase
journal, September 1991
- Boshart, Michael; Weih, Falk; Nichols, Mark
- Cell, Vol. 66, Issue 5
Subtractive hybridization cloning of a tissue-specific extinguisher: TSE1 encodes a regulatory subunit of protein kinase A
journal, September 1991
- Jones, Keith W.; Shapero, Michael H.; Chevrette, Mario
- Cell, Vol. 66, Issue 5
In vitro functional studies of naturally occurring pathogenic PRKAR1A mutations that are not subject to nonsense mRNA decay
journal, February 2008
- Greene, Elizabeth L.; Horvath, Anelia D.; Nesterova, Maria
- Human Mutation, Vol. 29, Issue 5
PRKAR1A Mutation Affecting cAMP-Mediated G Protein-Coupled Receptor Signaling in a Patient with Acrodysostosis and Hormone Resistance
journal, September 2012
- Nagasaki, Keisuke; Iida, Tomoko; Sato, Hidetoshi
- The Journal of Clinical Endocrinology & Metabolism, Vol. 97, Issue 9
Recurrent PRKAR1A Mutation in Acrodysostosis with Hormone Resistance
journal, June 2011
- Linglart, Agnès; Menguy, Christine; Couvineau, Alain
- New England Journal of Medicine, Vol. 364, Issue 23
Exome Sequencing Identifies PDE4D Mutations as Another Cause of Acrodysostosis
journal, April 2012
- Michot, Caroline; Le Goff, Carine; Goldenberg, Alice
- The American Journal of Human Genetics, Vol. 90, Issue 4
Acrodysostosis coinciding with pseudohypoparathyroidism and pseudo-pseudohypoparathyroidism
journal, January 1977
- Ablow, Rc; Hsia, Ye; Brandt, Ik
- American Journal of Roentgenology, Vol. 128, Issue 1
Crystal structures of the myristylated catalytic subunit of cAMP-dependent protein kinase reveal open and closed conformations
journal, October 1993
- Zheng, Jianhua; Knighton, Daniel R.; Taylor, Susan S.
- Protein Science, Vol. 2, Issue 10
The cAMP binding domain: An ancient signaling module
journal, December 2004
- Berman, H. M.; Ten Eyck, L. F.; Goodsell, D. S.
- Proceedings of the National Academy of Sciences, Vol. 102, Issue 1
In-vivo detection of binary PKA network interactions upon activation of endogenous GPCRs
journal, June 2015
- Röck, Ruth; Bachmann, Verena; Bhang, Hyo-eun C.
- Scientific Reports, Vol. 5, Issue 1
Quantification of dynamic protein complexes using Renilla luciferase fragment complementation applied to protein kinase A activities in vivo
journal, October 2007
- Stefan, E.; Aquin, S.; Berger, N.
- Proceedings of the National Academy of Sciences, Vol. 104, Issue 43
C Subunits Binding to the Protein Kinase A RIα Dimer Induce a Large Conformational Change
journal, April 2004
- Heller, William T.; Vigil, Dominico; Brown, Simon
- Journal of Biological Chemistry, Vol. 279, Issue 18
Arg-242 is necessary for allosteric coupling of cyclic AMP-binding sites A and B of RI subunit of cyclic AMP-dependent protein kinase.
journal, September 1994
- Symcox, M. M.; Cauthron, R. D.; Ogreid, D.
- Journal of Biological Chemistry, Vol. 269, Issue 37
PKA RIα Homodimer Structure Reveals an Intermolecular Interface with Implications for Cooperative cAMP Binding and Carney Complex Disease
journal, January 2014
- Bruystens, Jessica G. H.; Wu, Jian; Fortezzo, Audrey
- Structure, Vol. 22, Issue 1
Physiological inhibitors of the catalytic subunit of cAMP-dependent protein kinase: effect of magnesium-ATP on protein-protein interactions
journal, December 1993
- Herberg, Friedrich W.; Taylor, Susan S.
- Biochemistry, Vol. 32, Issue 50
Molecular Features of Product Release for the PKA Catalytic Cycle
journal, June 2014
- Bastidas, Adam C.; Wu, Jian; Taylor, Susan S.
- Biochemistry, Vol. 54, Issue 1
Contribution of Non-catalytic Core Residues to Activity and Regulation in Protein Kinase A
journal, January 2009
- Yang, Jie; Kennedy, Eileen J.; Wu, Jian
- Journal of Biological Chemistry, Vol. 284, Issue 10
Dissecting the cAMP-inducible allosteric switch in protein kinase A RIα
journal, April 2010
- Sjoberg, Timothy J.; Kornev, Alexandr P.; Taylor, Susan S.
- Protein Science, Vol. 19, Issue 6
Activation of type I cyclic AMP-dependent protein kinases with defective cyclic AMP-binding sites.
journal, February 1987
- Steinberg, R. A.; Russell, J. L.; Murphy, C. S.
- Journal of Biological Chemistry, Vol. 262, Issue 6
Hotspots for spontaneous and mutagen-induced lesions in regulatory subunit of cyclic AMP-dependent protein kinase in S49 mouse lymphoma cells
journal, November 1985
- Murphy, Caroline S.; Steinberg, Robert A.
- Somatic Cell and Molecular Genetics, Vol. 11, Issue 6
Dissecting cAMP Binding Domain A in the RIα Subunit of cAMP-dependent Protein Kinase
journal, October 1998
- Huang, Lily Jun-shen; Taylor, Susan S.
- Journal of Biological Chemistry, Vol. 273, Issue 41
Consequences of cAMP-Binding Site Mutations on the Structural Stability of the Type I Regulatory Subunit of cAMP-Dependent Protein Kinase
journal, November 2000
- Cànaves, Jaume M.; Leon, Darryl A.; Taylor, Susan S.
- Biochemistry, Vol. 39, Issue 49
Active Site Mutations Define the Pathway for the Cooperative Activation of cAMP-Dependent Protein Kinase †
journal, January 1996
- Herberg, Friedrich W.; Taylor, Susan S.; Dostmann, Wolfgang R. G.
- Biochemistry, Vol. 35, Issue 9
The kinetics of association of cyclic AMP to the two types of binding sites associated with protein kinase II from bovine myocardium
journal, July 1981
- Øgreid, Dagfinn; Døskeland, Stein Ove
- FEBS Letters, Vol. 129, Issue 2
The kinetics of the interaction between cyclic AMP and the regulatory moiety of protein kinase II: Evidence for interaction between the binding sites for cyclic AMP
journal, July 1981
- Øgreid, Dagfinn; Døskeland, Stein Ove
- FEBS Letters, Vol. 129, Issue 2
Probing the Multidomain Structure of the Type I Regulatory Subunit of cAMP-Dependent Protein Kinase Using Mutational Analysis: Role and Environment of Endogenous Tryptophans †
journal, May 2000
- Leon, Darryl A.; Canaves, Jaume M.; Taylor, Susan S.
- Biochemistry, Vol. 39, Issue 19
Using Markov State Models to Develop a Mechanistic Understanding of Protein Kinase A Regulatory Subunit RIα Activation in Response to cAMP Binding
journal, October 2014
- Boras, Britton W.; Kornev, Alexandr; Taylor, Susan S.
- Journal of Biological Chemistry, Vol. 289, Issue 43
PRKAR1A and PDE4D Mutations Cause Acrodysostosis but Two Distinct Syndromes with or without GPCR-Signaling Hormone Resistance
journal, December 2012
- Linglart, Agnès; Fryssira, Helena; Hiort, Olaf
- The Journal of Clinical Endocrinology & Metabolism, Vol. 97, Issue 12
Exome Sequencing Identifies PDE4D Mutations in Acrodysostosis
journal, April 2012
- Lee, Hane; Graham, John M.; Rimoin, David L.
- The American Journal of Human Genetics, Vol. 90, Issue 4
Novel mutations of the PRKAR1A gene in patients with acrodysostosis: Novel mutations of PRKAR1A
journal, February 2013
- Muhn, F.; Klopocki, E.; Graul-Neumann, L.
- Clinical Genetics, Vol. 84, Issue 6
Screening of PRKAR1A and PDE4D in a Large Italian Series of Patients Clinically Diagnosed With Albright Hereditary Osteodystrophy and/or Pseudohypoparathyroidism: SCREENING FOR PRKAR1A AND PDE4D GENETIC DEFECTS
journal, February 2016
- Elli, Francesca Marta; Bordogna, Paolo; de Sanctis, Luisa
- Journal of Bone and Mineral Research, Vol. 31, Issue 6
Regulatory subunit of protein kinase A: structure of deletion mutant with cAMP binding domains
journal, August 1995
- Su, Y.; Dostmann, W.; Herberg, F.
- Science, Vol. 269, Issue 5225
Crystal Structures of RIα Subunit of Cyclic Adenosine 5‘-Monophosphate (cAMP)-Dependent Protein Kinase Complexed with ( R p )-Adenosine 3‘,5‘-Cyclic Monophosphothioate and ( S p )-Adenosine 3‘,5‘-Cyclic Monophosphothioate, the Phosphothioate Analogues of cAMP † , ‡
journal, June 2004
- Wu, Jian; Jones, John M.; Xuong, Nguyen-Huu
- Biochemistry, Vol. 43, Issue 21
Backbone Flexibility of Five Sites on the Catalytic Subunit of cAMP-Dependent Protein Kinase in the Open and Closed Conformations
journal, September 1998
- Gangal, Milind; Cox, Sarah; Lew, John
- Biochemistry, Vol. 37, Issue 39
Assay Principle for Modulators of Protein−Protein Interactions and Its Application to Non-ATP-Competitive Ligands Targeting Protein Kinase A
journal, December 2006
- Saldanha, S. Adrian; Kaler, Gregory; Cottam, Howard B.
- Analytical Chemistry, Vol. 78, Issue 24
[20] Processing of X-ray diffraction data collected in oscillation mode
book, January 1997
- Otwinowski, Zbyszek; Minor, Wladek
- Macromolecular Crystallography Part A, 307-326
PROCHECK: a program to check the stereochemical quality of protein structures
journal, April 1993
- Laskowski, R. A.; MacArthur, M. W.; Moss, D. S.
- Journal of Applied Crystallography, Vol. 26, Issue 2
Determination of the regularization parameter in indirect-transform methods using perceptual criteria
journal, August 1992
- Svergun, D. I.
- Journal of Applied Crystallography, Vol. 25, Issue 4, p. 495-503
Conformational Differences Among Solution Structures of the Type Iα, IIα and IIβ Protein Kinase A Regulatory Subunit Homodimers: Role of the Linker Regions
journal, April 2004
- Vigil, Dominico; Blumenthal, Donald K.; Heller, William T.
- Journal of Molecular Biology, Vol. 337, Issue 5
Works referencing / citing this record:
Switching of the folding-energy landscape governs the allosteric activation of protein kinase A
journal, July 2018
- England, Jeneffer P.; Hao, Yuxin; Bai, Lihui
- Proceedings of the National Academy of Sciences, Vol. 115, Issue 32
Cardiac function modulation depends on the A‐kinase anchoring protein complex
journal, August 2019
- Zhu, Yan‐Rong; Jiang, Xiao‐Xin; Zheng, Yaguo
- Journal of Cellular and Molecular Medicine, Vol. 23, Issue 11
Cardiac function modulation depends on the A‐kinase anchoring protein complex
journal, August 2019
- Zhu, Yan‐Rong; Jiang, Xiao‐Xin; Zheng, Yaguo
- Journal of Cellular and Molecular Medicine, Vol. 23, Issue 11