Phosphorylation and Methylation of Proteasomal Proteins of the Haloarcheon Haloferax volcanii
Abstract
Proteasomes are composed of 20S core particles (CPs) of - and -type subunits that associate with regulatory particle AAA ATPases such as the proteasome-activating nucleotidase (PAN) complexes of archaea. In this study, the roles and additional sites of post-translational modification of proteasomes were investigated using the archaeonHaloferax volcaniias a model. Indicative of phosphorylation, phosphatase-sensitive isoforms of and were detected by 2-DE immunoblot. To map these and other potential sites of post-translational modification, proteasomes were purified and analyzed by tandem mass spectrometry (MS/MS). Using this approach, several phosphosites were mapped including Thr147, 2 Thr13/Ser14 and PAN-A Ser340. Multiple methylation sites were also mapped to , thus, revealing a new type of proteasomal modification. Probing the biological role of and PAN-A phosphorylation by site-directed mutagenesis revealed dominant negative phenotypes for cell viability and/or pigmentation for variants including Thr147Ala, Thr158Ala and Ser58Ala. AnH. volcaniiRio1p Ser/Thr kinase homolog was purified and shown to catalyze autophosphorylation and phosphotransfer to . The variants in Thr and Ser residues that displayed dominant negative phenotypes were significantly reduced in their ability to accept phosphoryl groups from Rio1p, thus, providing an important link between cell physiology and proteasomal phosphorylation.
- Authors:
-
- Department of Microbiology and Cell Science, University of Florida, Gainesville, FL 32611, USA
- Publication Date:
- Research Org.:
- Univ. of Florida, Gainesville, FL (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC), Biological and Environmental Research (BER). Biological Systems Science Division
- OSTI Identifier:
- 1198438
- Alternate Identifier(s):
- OSTI ID: 1626200
- Grant/Contract Number:
- FG02-05ER15650
- Resource Type:
- Published Article
- Journal Name:
- Archaea
- Additional Journal Information:
- Journal Name: Archaea Journal Volume: 2010; Journal ID: ISSN 1472-3646
- Publisher:
- Hindawi
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; Microbiology
Citation Formats
Humbard, Matthew A., Reuter, Christopher J., Zuobi-Hasona, Kheir, Zhou, Guangyin, and Maupin-Furlow, Julie A. Phosphorylation and Methylation of Proteasomal Proteins of the Haloarcheon Haloferax volcanii. United States: N. p., 2010.
Web. doi:10.1155/2010/481725.
Humbard, Matthew A., Reuter, Christopher J., Zuobi-Hasona, Kheir, Zhou, Guangyin, & Maupin-Furlow, Julie A. Phosphorylation and Methylation of Proteasomal Proteins of the Haloarcheon Haloferax volcanii. United States. https://doi.org/10.1155/2010/481725
Humbard, Matthew A., Reuter, Christopher J., Zuobi-Hasona, Kheir, Zhou, Guangyin, and Maupin-Furlow, Julie A. Fri .
"Phosphorylation and Methylation of Proteasomal Proteins of the Haloarcheon Haloferax volcanii". United States. https://doi.org/10.1155/2010/481725.
@article{osti_1198438,
title = {Phosphorylation and Methylation of Proteasomal Proteins of the Haloarcheon Haloferax volcanii},
author = {Humbard, Matthew A. and Reuter, Christopher J. and Zuobi-Hasona, Kheir and Zhou, Guangyin and Maupin-Furlow, Julie A.},
abstractNote = {Proteasomes are composed of 20S core particles (CPs) ofα- andβ-type subunits that associate with regulatory particle AAA ATPases such as the proteasome-activating nucleotidase (PAN) complexes of archaea. In this study, the roles and additional sites of post-translational modification of proteasomes were investigated using the archaeonHaloferax volcaniias a model. Indicative of phosphorylation, phosphatase-sensitive isoforms ofα1andα2were detected by 2-DE immunoblot. To map these and other potential sites of post-translational modification, proteasomes were purified and analyzed by tandem mass spectrometry (MS/MS). Using this approach, several phosphosites were mapped includingα1Thr147,α2 Thr13/Ser14 and PAN-A Ser340. Multiple methylation sites were also mapped toα1, thus, revealing a new type of proteasomal modification. Probing the biological role ofα1and PAN-A phosphorylation by site-directed mutagenesis revealed dominant negative phenotypes for cell viability and/or pigmentation forα1variants including Thr147Ala, Thr158Ala and Ser58Ala. AnH. volcaniiRio1p Ser/Thr kinase homolog was purified and shown to catalyze autophosphorylation and phosphotransfer toα1. Theα1variants in Thr and Ser residues that displayed dominant negative phenotypes were significantly reduced in their ability to accept phosphoryl groups from Rio1p, thus, providing an important link between cell physiology and proteasomal phosphorylation.},
doi = {10.1155/2010/481725},
journal = {Archaea},
number = ,
volume = 2010,
place = {United States},
year = {2010},
month = {1}
}
https://doi.org/10.1155/2010/481725
Web of Science
Works referenced in this record:
PRS3 encoding an essential subunit of yeast proteasomes homologous to mammalian proteasome subunit C5
journal, January 1992
- Lee, Do Hee; Tanaka, Keiji; Tamura, Tomohiro
- Biochemical and Biophysical Research Communications, Vol. 182, Issue 2
Molecular cloning of an essential yeast gene encoding a proteasomal subunit
journal, March 1992
- Georgatsou, Elena; Georgakopoulos, Tassos; Thireos, George
- FEBS Letters, Vol. 299, Issue 1
Proteasomal Components Required for Cell Growth and Stress Responses in the Haloarchaeon Haloferax volcanii
journal, October 2008
- Zhou, G.; Kowalczyk, D.; Humbard, M. A.
- Journal of Bacteriology, Vol. 190, Issue 24
Eukaryotic 20S proteasome catalytic subunit propeptides prevent active site inactivation by N-terminal acetylation and promote particle assembly
journal, July 1999
- Arendt, C. S.
- The EMBO Journal, Vol. 18, Issue 13
N-Terminal modifications of the 19S regulatory particle subunits of the yeast proteasome
journal, January 2003
- Kimura, Yayoi; Saeki, Yasushi; Yokosawa, Hideyoshi
- Archives of Biochemistry and Biophysics, Vol. 409, Issue 2
N α -Acetylation and Proteolytic Activity of the Yeast 20 S Proteasome
journal, February 2000
- Kimura, Yayoi; Takaoka, Motoko; Tanaka, Sono
- Journal of Biological Chemistry, Vol. 275, Issue 7
The NH 2 -terminal residues of rat liver proteasome (multicatalytic proteinase complex) subunits, C2, C3 and C8, are N α-acetylated
journal, April 1990
- Tokunaga, Fuminori; Aruga, Rie; Iwanaga, Sadaaki
- FEBS Letters, Vol. 263, Issue 2
Hyperphosphorylation of rat liver proteasome subunits: the effects of ethanol and okadaic acid are compared
journal, June 2004
- Bardaggorce, F.
- Life Sciences, Vol. 75, Issue 5
Phosphorylation of 20S proteasome alpha subunit C8 (alpha7) stabilizes the 26S proteasome and plays a role in the regulation of proteasome complexes by gamma-interferon
journal, February 2004
- Bose, Suchira; Stratford, Fiona L. L.; Broadfoot, Kerry I.
- Biochemical Journal, Vol. 378, Issue 1
γ-Interferon decreases the level of 26 S proteasomes and changes the pattern of phosphorylation
journal, January 2001
- Bose, Suchira; Brooks, Paul; Mason, Grant G. F.
- Biochemical Journal, Vol. 353, Issue 2
Phosphorylation of C8 and C9 Subunits of the Multicatalytic Proteinase by Casein Kinase II and Identification of the C8 Phosphorylation Sites by Direct Mutagenesis †
journal, January 1996
- Castaño, José G.; Mahillo, Esther; Arizti, Paz
- Biochemistry, Vol. 35, Issue 12
Mapping and Structural Dissection of Human 20 S Proteasome Using Proteomic Approaches
journal, August 2002
- Claverol, Stephane; Burlet-Schiltz, Odile; Girbal-Neuhauser, Elisabeth
- Molecular & Cellular Proteomics, Vol. 1, Issue 8
In vivo and in vitro phosphorylation of Candida albicans 20S proteasome
journal, August 2002
- Fernández Murray, Pedro; Pardo, Patricia S.; Zelada, Alicia M.
- Archives of Biochemistry and Biophysics, Vol. 404, Issue 1
Identification of a protein kinase which phosphorylates a subunit of the 26S proteasome and changes in its activity during meiotic cell cycle in goldfish oocytes
journal, February 2005
- Horiguchi, Ryo; Yoshikuni, Michiyasu; Tokumoto, Mika
- Cellular Signalling, Vol. 17, Issue 2
Identification of three phosphorylation sites in the α7 subunit of the yeast 20S proteasome in vivo using mass spectrometry
journal, November 2004
- Iwafune, Yuko; Kawasaki, Hiroshi; Hirano, Hisashi
- Archives of Biochemistry and Biophysics, Vol. 431, Issue 1
Phosphorylation of Proteasomes in Mammalian Cells. Identification of Two Phosphorylated Subunits and the Effect of Phosphorylation on Activity
journal, June 1996
- Mason, Grant G. F.; Hendil, Klavs B.; Rivett, A. Jennifer
- European Journal of Biochemistry, Vol. 238, Issue 2
Phosphorylation of ATPase subunits of the 26S proteasome
journal, July 1998
- Mason, Grant G. F.; Murray, Rachael Z.; Pappin, Daryl
- FEBS Letters, Vol. 430, Issue 3
In Vivoandin VitroPhosphorylation of the α7/PRS1 Subunit ofSaccharomyces cerevisiae20 S Proteasome:In VitroPhosphorylation by Protein Kinase CK2 Is Absolutely Dependent on Polylysine
journal, January 1998
- Pardo, Patricia S.; Murray, Pedro Fernández; Walz, Katherina
- Archives of Biochemistry and Biophysics, Vol. 349, Issue 2
Regulation of proteasome complexes by γ-interferon and phosphorylation
journal, March 2001
- Jennifer Rivett, A.; Bose, Suchira; Brooks, Paul
- Biochimie, Vol. 83, Issue 3-4
Assembly of the 26S Proteasome Is Regulated by Phosphorylation of the p45/Rpt6 ATPase Subunit †
journal, January 2001
- Satoh, Kazuki; Sasajima, Hitoshi; Nyoumura, Ken-ichi
- Biochemistry, Vol. 40, Issue 2
Two proteins, a goldfish 20S proteasome subunit and the protein interacting with 26S proteasome, change in the meiotic cell cycle: Changes in 26S proteasome during meiotic cell cycle
journal, January 2000
- Tokumoto, Mika; Horiguchi, Ryo; Nagahama, Yoshitaka
- European Journal of Biochemistry, Vol. 267, Issue 1
Phosphorylation of the C2 subunit of the proteasome in rice ( Oryza sativa L.)
journal, February 1997
- Umeda, Masaaki; Manabe, Yuzuki; Uchimiya, Hirofumi
- FEBS Letters, Vol. 403, Issue 3
Proteasome Function Is Regulated by Cyclic AMP-dependent Protein Kinase through Phosphorylation of Rpt6
journal, August 2007
- Zhang, Fengxue; Hu, Yong; Huang, Ping
- Journal of Biological Chemistry, Vol. 282, Issue 31
20 S Proteasome from Saccharomyces cerevisiae Is Responsive to Redox Modifications and IsS-Glutathionylated
journal, January 2003
- Demasi, Marilene; Silva, Gustavo Monteiro; Netto, Luis Eduardo Soares
- Journal of Biological Chemistry, Vol. 278, Issue 1
A novel interaction between N‐myristoylation and the 26S proteasome during cell morphogenesis
journal, January 2007
- Lee, Soo Chan; Shaw, Brian D.
- Molecular Microbiology, Vol. 63, Issue 4
O-GlcNAc modification: a nutritional sensor that modulates proteasome function
journal, May 2004
- Zachara, Natasha E.; Hart, Gerald W.
- Trends in Cell Biology, Vol. 14, Issue 5
Proteasome from Thermoplasma acidophilum: a threonine protease
journal, April 1995
- Seemuller, E.; Lupas, A.; Stock, D.
- Science, Vol. 268, Issue 5210
Regulation of the Proteasome by Neuronal Activity and Calcium/Calmodulin-dependent Protein Kinase II
journal, September 2009
- Djakovic, Stevan N.; Schwarz, Lindsay A.; Barylko, Barbara
- Journal of Biological Chemistry, Vol. 284, Issue 39
Halophilic 20S Proteasomes of the ArchaeonHaloferax volcanii: Purification, Characterization, and Gene Sequence Analysis
journal, September 1999
- Wilson, Heather L.; Aldrich, Henry C.; Maupin-Furlow, Julie
- Journal of Bacteriology, Vol. 181, Issue 18
Posttranslational Modification of the 20S Proteasomal Proteins of the Archaeon Haloferax volcanii
journal, September 2006
- Humbard, Matthew A.; Stevens, Stanley M.; Maupin-Furlow, Julie A.
- Journal of Bacteriology, Vol. 188, Issue 21
Subunit Topology of Two 20S Proteasomes from Haloferax volcanii
journal, January 2003
- Kaczowka, Steven J.; Maupin-Furlow, Julie A.
- Journal of Bacteriology, Vol. 185, Issue 1
Differential Regulation of the PanA and PanB Proteasome-Activating Nucleotidase and 20S Proteasomal Proteins of the Haloarchaeon Haloferax volcanii
journal, November 2004
- Reuter, Christopher J.; Kaczowka, Steven J.; Maupin-Furlow, Julie A.
- Journal of Bacteriology, Vol. 186, Issue 22
The N-Terminal Penultimate Residue of 20S Proteasome 1 Influences its N Acetylation and Protein Levels as Well as Growth Rate and Stress Responses of Haloferax volcanii
journal, April 2009
- Humbard, M. A.; Zhou, G.; Maupin-Furlow, J. A.
- Journal of Bacteriology, Vol. 191, Issue 12
Trizol-based method for sample preparation and isoelectric focusing of halophilic proteins
journal, April 2006
- Kirkland, P. A.; Busby, Jennifer; Stevens, Stan
- Analytical Biochemistry, Vol. 351, Issue 2
Fluorimetric assays for cathepsin B and cathepsin H with methylcoumarylamide substrates
journal, June 1980
- Barrett, A. J.
- Biochemical Journal, Vol. 187, Issue 3
Sequence and structure-based prediction of eukaryotic protein phosphorylation sites
journal, December 1999
- Blom, Nikolaj; Gammeltoft, Steen; Brunak, Søren
- Journal of Molecular Biology, Vol. 294, Issue 5
Cloning and characterization of the Bacillus subtilis prkA gene encoding a novel serine protein kinase
journal, January 1996
- Fischer, Cécile; Geourjon, Christophe; Bourson, Claude
- Gene, Vol. 168, Issue 1
Autophosphorylation of Archaeoglobus fulgidus Rio2 and crystal structures of its nucleotide-metal ion complexes: Nucleotide-metal ion complexes of Rio2
journal, May 2005
- LaRonde-LeBlanc, Nicole; Guszczynski, Tad; Copeland, Terry
- FEBS Journal, Vol. 272, Issue 11
A Family Portrait of the RIO Kinases
journal, September 2005
- LaRonde-LeBlanc, Nicole; Wlodawer, Alexander
- Journal of Biological Chemistry, Vol. 280, Issue 45
Transcriptional linkage of Haloferax volcanii proteasomal genes with non-proteasomal gene neighbours including RNase P, MOSC domain and SAM-methyltransferase homologues
journal, September 2007
- Gil, Malgorzata A.; Sherwood, Katherine E.; Maupin-Furlow, Julie A.
- Microbiology, Vol. 153, Issue 9
Archaeal Eukaryote-Like Serine/Threonine Protein Kinase Interacts with and Phosphorylates a Forkhead-Associated-Domain-Containing Protein
journal, January 2010
- Wang, Bin; Yang, Shifan; Zhang, Lei
- Journal of Bacteriology, Vol. 192, Issue 7
The regulatory crosstalk between kinases and proteases in cancer
journal, March 2010
- López-Otín, Carlos; Hunter, Tony
- Nature Reviews Cancer, Vol. 10, Issue 4
Protein Quality Control as a Strategy for Cellular Regulation: Lessons from Ubiquitin-Mediated Regulation of the Sterol Pathway
journal, April 2009
- Hampton, Randolph Y.; Garza, Renee M.
- Chemical Reviews, Vol. 109, Issue 4
A RING-finger protein regulates carotenogenesis via proteolysis-independent ubiquitylation of a White Collar-1-like activator
journal, November 2008
- Silva, Fátima; Navarro, Eusebio; Peñaranda, Ascensión
- Molecular Microbiology
Primary structure and functional analysis of the soluble transducer protein HtrXI in the archaeon Halobacterium salinarium.
journal, January 1997
- Brooun, A.; Zhang, W.; Alam, M.
- Journal of bacteriology, Vol. 179, Issue 9
The lifetime of photosensory signals in Halobacterium halobium and its dependence on protein methylation
journal, April 1990
- Hildebrand, Eilo; Schimz, Angelika
- Biochimica et Biophysica Acta (BBA) - Molecular Cell Research, Vol. 1052, Issue 1
Sensory Rhodopsin II Transducer HtrII Is Also Responsible for Serine Chemotaxis in the ArchaeonHalobacterium salinarum
journal, March 1998
- Hou, Shaobin; Brooun, Alexei; Suk Yu, Hyung
- Journal of Bacteriology, Vol. 180, Issue 6