Dynamic regulation of a metabolic multi-enzyme complex by protein kinase CK2

Songon An, Minjoung Kyoung, Jasmina J. Allen, Kevan M. Shokat, Stephen Benkovic

Research output: Contribution to journalArticle

42 Citations (Scopus)

Abstract

The reversible association and dissociation of a metabolic multi-enzyme complex participating in de novo purine biosynthesis, the purinosome, was demonstrated in live cells to respond to the levels of purine nucleotides in the culture media. We also took advantage of in vitro proteomic scale studies of cellular substrates of human protein kinases (e.g. casein kinase II (CK2) and Akt), that implicated several de novo purine biosynthetic enzymes as kinase substrates. Here, we successfully identified that purinosome formation in vivo was significantly promoted in HeLa cells by the addition of small-molecule CK2-specific inhibitors (i.e. 4,5,6,7-tetrabromo-1H-benzimidazole, 2-dimethylamino-4,5,6,7-tetrabromo-1H-benzimidazole, tetrabromocinammic acid, 4,4′,5,5′,6,6′-hexahydroxydiphenic acid 2,2′,6,6′- dilactone (ellagic acid) as well as by silencing the endogenous human CK2αcatalytic subunit with small interfering RNA. However, 4,5,6,7-tetrabromobenzotriazole, another CK2-specific inhibitor, triggered the dissociation of purinosome clusters in HeLa cells. Although the mechanism by which 4,5,6,7-tetrabromobenzotriazole affects purinosome clustering is not clear, we were capable of chemically reversing purinosome formation in cells by the sequential addition of two CK2 inhibitors. Collectively, we provide compelling cellular evidence that CK2-mediated pathways reversibly regulate purinosome assembly, and thus the purinosome may be one of the ultimate targets of kinase inhibitors.

Original languageEnglish (US)
Pages (from-to)11093-11099
Number of pages7
JournalJournal of Biological Chemistry
Volume285
Issue number15
DOIs
StatePublished - Apr 9 2010

Fingerprint

Casein Kinase II
HeLa Cells
Phosphotransferases
Ellagic Acid
Purine Nucleotides
Acids
Biosynthesis
Substrates
Enzymes
Proteomics
Protein Kinases
Small Interfering RNA
Cluster Analysis
Culture Media
Association reactions
Molecules
purine
4,5,6,7-tetrabromobenzotriazole
Multienzyme Complexes
2-dimethylamino-4,5,6,7-tetrabromo-1H-benzimidazole

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

An, S., Kyoung, M., Allen, J. J., Shokat, K. M., & Benkovic, S. (2010). Dynamic regulation of a metabolic multi-enzyme complex by protein kinase CK2. Journal of Biological Chemistry, 285(15), 11093-11099. https://doi.org/10.1074/jbc.M110.101139
An, Songon ; Kyoung, Minjoung ; Allen, Jasmina J. ; Shokat, Kevan M. ; Benkovic, Stephen. / Dynamic regulation of a metabolic multi-enzyme complex by protein kinase CK2. In: Journal of Biological Chemistry. 2010 ; Vol. 285, No. 15. pp. 11093-11099.
@article{928265106f4c4809abe1df39a8b58d70,
title = "Dynamic regulation of a metabolic multi-enzyme complex by protein kinase CK2",
abstract = "The reversible association and dissociation of a metabolic multi-enzyme complex participating in de novo purine biosynthesis, the purinosome, was demonstrated in live cells to respond to the levels of purine nucleotides in the culture media. We also took advantage of in vitro proteomic scale studies of cellular substrates of human protein kinases (e.g. casein kinase II (CK2) and Akt), that implicated several de novo purine biosynthetic enzymes as kinase substrates. Here, we successfully identified that purinosome formation in vivo was significantly promoted in HeLa cells by the addition of small-molecule CK2-specific inhibitors (i.e. 4,5,6,7-tetrabromo-1H-benzimidazole, 2-dimethylamino-4,5,6,7-tetrabromo-1H-benzimidazole, tetrabromocinammic acid, 4,4′,5,5′,6,6′-hexahydroxydiphenic acid 2,2′,6,6′- dilactone (ellagic acid) as well as by silencing the endogenous human CK2αcatalytic subunit with small interfering RNA. However, 4,5,6,7-tetrabromobenzotriazole, another CK2-specific inhibitor, triggered the dissociation of purinosome clusters in HeLa cells. Although the mechanism by which 4,5,6,7-tetrabromobenzotriazole affects purinosome clustering is not clear, we were capable of chemically reversing purinosome formation in cells by the sequential addition of two CK2 inhibitors. Collectively, we provide compelling cellular evidence that CK2-mediated pathways reversibly regulate purinosome assembly, and thus the purinosome may be one of the ultimate targets of kinase inhibitors.",
author = "Songon An and Minjoung Kyoung and Allen, {Jasmina J.} and Shokat, {Kevan M.} and Stephen Benkovic",
year = "2010",
month = "4",
day = "9",
doi = "10.1074/jbc.M110.101139",
language = "English (US)",
volume = "285",
pages = "11093--11099",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "15",

}

An, S, Kyoung, M, Allen, JJ, Shokat, KM & Benkovic, S 2010, 'Dynamic regulation of a metabolic multi-enzyme complex by protein kinase CK2', Journal of Biological Chemistry, vol. 285, no. 15, pp. 11093-11099. https://doi.org/10.1074/jbc.M110.101139

Dynamic regulation of a metabolic multi-enzyme complex by protein kinase CK2. / An, Songon; Kyoung, Minjoung; Allen, Jasmina J.; Shokat, Kevan M.; Benkovic, Stephen.

In: Journal of Biological Chemistry, Vol. 285, No. 15, 09.04.2010, p. 11093-11099.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Dynamic regulation of a metabolic multi-enzyme complex by protein kinase CK2

AU - An, Songon

AU - Kyoung, Minjoung

AU - Allen, Jasmina J.

AU - Shokat, Kevan M.

AU - Benkovic, Stephen

PY - 2010/4/9

Y1 - 2010/4/9

N2 - The reversible association and dissociation of a metabolic multi-enzyme complex participating in de novo purine biosynthesis, the purinosome, was demonstrated in live cells to respond to the levels of purine nucleotides in the culture media. We also took advantage of in vitro proteomic scale studies of cellular substrates of human protein kinases (e.g. casein kinase II (CK2) and Akt), that implicated several de novo purine biosynthetic enzymes as kinase substrates. Here, we successfully identified that purinosome formation in vivo was significantly promoted in HeLa cells by the addition of small-molecule CK2-specific inhibitors (i.e. 4,5,6,7-tetrabromo-1H-benzimidazole, 2-dimethylamino-4,5,6,7-tetrabromo-1H-benzimidazole, tetrabromocinammic acid, 4,4′,5,5′,6,6′-hexahydroxydiphenic acid 2,2′,6,6′- dilactone (ellagic acid) as well as by silencing the endogenous human CK2αcatalytic subunit with small interfering RNA. However, 4,5,6,7-tetrabromobenzotriazole, another CK2-specific inhibitor, triggered the dissociation of purinosome clusters in HeLa cells. Although the mechanism by which 4,5,6,7-tetrabromobenzotriazole affects purinosome clustering is not clear, we were capable of chemically reversing purinosome formation in cells by the sequential addition of two CK2 inhibitors. Collectively, we provide compelling cellular evidence that CK2-mediated pathways reversibly regulate purinosome assembly, and thus the purinosome may be one of the ultimate targets of kinase inhibitors.

AB - The reversible association and dissociation of a metabolic multi-enzyme complex participating in de novo purine biosynthesis, the purinosome, was demonstrated in live cells to respond to the levels of purine nucleotides in the culture media. We also took advantage of in vitro proteomic scale studies of cellular substrates of human protein kinases (e.g. casein kinase II (CK2) and Akt), that implicated several de novo purine biosynthetic enzymes as kinase substrates. Here, we successfully identified that purinosome formation in vivo was significantly promoted in HeLa cells by the addition of small-molecule CK2-specific inhibitors (i.e. 4,5,6,7-tetrabromo-1H-benzimidazole, 2-dimethylamino-4,5,6,7-tetrabromo-1H-benzimidazole, tetrabromocinammic acid, 4,4′,5,5′,6,6′-hexahydroxydiphenic acid 2,2′,6,6′- dilactone (ellagic acid) as well as by silencing the endogenous human CK2αcatalytic subunit with small interfering RNA. However, 4,5,6,7-tetrabromobenzotriazole, another CK2-specific inhibitor, triggered the dissociation of purinosome clusters in HeLa cells. Although the mechanism by which 4,5,6,7-tetrabromobenzotriazole affects purinosome clustering is not clear, we were capable of chemically reversing purinosome formation in cells by the sequential addition of two CK2 inhibitors. Collectively, we provide compelling cellular evidence that CK2-mediated pathways reversibly regulate purinosome assembly, and thus the purinosome may be one of the ultimate targets of kinase inhibitors.

UR - http://www.scopus.com/inward/record.url?scp=77951236102&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=77951236102&partnerID=8YFLogxK

U2 - 10.1074/jbc.M110.101139

DO - 10.1074/jbc.M110.101139

M3 - Article

VL - 285

SP - 11093

EP - 11099

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 15

ER -