PKA catalytic subunit mutations in adrenocortical Cushing's adenoma impair association with the regulatory subunit

Davide Calebiro; Cristina Ronchi; Annette Hannawacker; Sandra Lyga; Kerstin Bathon; Ulrike Zabel; Felix Beuschlein; Martin Reincke; Kristina Lorenz; Bruno Allolio; Caroline Kisker; Martin Fassnacht; Martin J. Lohse

doi:10.1038/ncomms6680

PKA catalytic subunit mutations in adrenocortical Cushing's adenoma impair association with the regulatory subunit

Davide Calebiro^*, Cristina Ronchi, Annette Hannawacker, Sandra Lyga, Kerstin Bathon, Ulrike Zabel, Felix Beuschlein, Martin Reincke, Kristina Lorenz, Bruno Allolio, Caroline Kisker, Martin Fassnacht, Martin J. Lohse

^*Corresponding author for this work

Metabolism and Systems Research

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

41 Citations (Scopus)

125 Downloads (Pure)

Abstract

We recently identified a high prevalence of mutations affecting the catalytic (Cα) subunit of protein kinase A (PKA) in cortisol-secreting adrenocortical adenomas. The two identified mutations (Leu206Arg and Leu199-Cys200insTrp) are associated with increased PKA catalytic activity, but the underlying mechanisms are highly controversial. Here we utilize a combination of biochemical and optical assays, including fluorescence resonance energy transfer in living cells, to analyze the consequences of the two mutations with respect to the formation of the PKA holoenzyme and its regulation by cAMP. Our results indicate that neither mutant can form a stable PKA complex, due to the location of the mutations at the interface between the catalytic and the regulatory subunits. We conclude that the two mutations cause high basal catalytic activity and lack of regulation by cAMP through interference of complex formation between the regulatory and the catalytic subunits of PKA.

Original language	English
Article number	5680
Number of pages	7
Journal	Nature Communications
Volume	5
DOIs	https://doi.org/10.1038/ncomms6680
Publication status	Published - 5 Dec 2014

Keywords

Adrenal tumours
Biochemistry
Mutation

ASJC Scopus subject areas

Chemistry(all)
Biochemistry, Genetics and Molecular Biology(all)
Physics and Astronomy(all)

Access to Document

10.1038/ncomms6680Licence: Creative Commons: Attribution (CC BY)

Calebiro_PKA_catalytic_subunit_mutations_Nature_Communications
Published as above, final version of record available at doi:10.1038/ncomms6680. Checked 17/5/18.
Final published version, 892 KBLicence: Creative Commons: Attribution (CC BY)

Cite this

Calebiro, D., Ronchi, C., Hannawacker, A., Lyga, S., Bathon, K., Zabel, U., Beuschlein, F., Reincke, M., Lorenz, K., Allolio, B., Kisker, C., Fassnacht, M., & Lohse, M. J. (2014). PKA catalytic subunit mutations in adrenocortical Cushing's adenoma impair association with the regulatory subunit. Nature Communications, 5, Article 5680. https://doi.org/10.1038/ncomms6680

@article{c08eb5edafe94468a125443a16c502eb,

title = "PKA catalytic subunit mutations in adrenocortical Cushing's adenoma impair association with the regulatory subunit",

abstract = "We recently identified a high prevalence of mutations affecting the catalytic (Cα) subunit of protein kinase A (PKA) in cortisol-secreting adrenocortical adenomas. The two identified mutations (Leu206Arg and Leu199-Cys200insTrp) are associated with increased PKA catalytic activity, but the underlying mechanisms are highly controversial. Here we utilize a combination of biochemical and optical assays, including fluorescence resonance energy transfer in living cells, to analyze the consequences of the two mutations with respect to the formation of the PKA holoenzyme and its regulation by cAMP. Our results indicate that neither mutant can form a stable PKA complex, due to the location of the mutations at the interface between the catalytic and the regulatory subunits. We conclude that the two mutations cause high basal catalytic activity and lack of regulation by cAMP through interference of complex formation between the regulatory and the catalytic subunits of PKA.",

keywords = "Adrenal tumours, Biochemistry, Mutation",

author = "Davide Calebiro and Cristina Ronchi and Annette Hannawacker and Sandra Lyga and Kerstin Bathon and Ulrike Zabel and Felix Beuschlein and Martin Reincke and Kristina Lorenz and Bruno Allolio and Caroline Kisker and Martin Fassnacht and Lohse, {Martin J.}",

year = "2014",

month = dec,

day = "5",

doi = "10.1038/ncomms6680",

language = "English",

volume = "5",

journal = "Nature Communications",

issn = "2041-1723",

publisher = "Springer",

}

TY - JOUR

T1 - PKA catalytic subunit mutations in adrenocortical Cushing's adenoma impair association with the regulatory subunit

AU - Calebiro, Davide

AU - Ronchi, Cristina

AU - Hannawacker, Annette

AU - Lyga, Sandra

AU - Bathon, Kerstin

AU - Zabel, Ulrike

AU - Beuschlein, Felix

AU - Reincke, Martin

AU - Lorenz, Kristina

AU - Allolio, Bruno

AU - Kisker, Caroline

AU - Fassnacht, Martin

AU - Lohse, Martin J.

PY - 2014/12/5

Y1 - 2014/12/5

N2 - We recently identified a high prevalence of mutations affecting the catalytic (Cα) subunit of protein kinase A (PKA) in cortisol-secreting adrenocortical adenomas. The two identified mutations (Leu206Arg and Leu199-Cys200insTrp) are associated with increased PKA catalytic activity, but the underlying mechanisms are highly controversial. Here we utilize a combination of biochemical and optical assays, including fluorescence resonance energy transfer in living cells, to analyze the consequences of the two mutations with respect to the formation of the PKA holoenzyme and its regulation by cAMP. Our results indicate that neither mutant can form a stable PKA complex, due to the location of the mutations at the interface between the catalytic and the regulatory subunits. We conclude that the two mutations cause high basal catalytic activity and lack of regulation by cAMP through interference of complex formation between the regulatory and the catalytic subunits of PKA.

AB - We recently identified a high prevalence of mutations affecting the catalytic (Cα) subunit of protein kinase A (PKA) in cortisol-secreting adrenocortical adenomas. The two identified mutations (Leu206Arg and Leu199-Cys200insTrp) are associated with increased PKA catalytic activity, but the underlying mechanisms are highly controversial. Here we utilize a combination of biochemical and optical assays, including fluorescence resonance energy transfer in living cells, to analyze the consequences of the two mutations with respect to the formation of the PKA holoenzyme and its regulation by cAMP. Our results indicate that neither mutant can form a stable PKA complex, due to the location of the mutations at the interface between the catalytic and the regulatory subunits. We conclude that the two mutations cause high basal catalytic activity and lack of regulation by cAMP through interference of complex formation between the regulatory and the catalytic subunits of PKA.

KW - Adrenal tumours

KW - Biochemistry

KW - Mutation

UR - http://www.scopus.com/inward/record.url?scp=84923279889&partnerID=8YFLogxK

U2 - 10.1038/ncomms6680

DO - 10.1038/ncomms6680

M3 - Article

C2 - 25477193

AN - SCOPUS:84923279889

SN - 2041-1723

VL - 5

JO - Nature Communications

JF - Nature Communications

M1 - 5680

ER -

PKA catalytic subunit mutations in adrenocortical Cushing's adenoma impair association with the regulatory subunit

Abstract

Keywords

ASJC Scopus subject areas

Access to Document

Fingerprint

Cite this