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Title: SDS22 selectively recognizes and traps metal-deficient inactive PP1

Abstract

The metalloenzyme protein phosphatase 1 (PP1), which is responsible for ≥50% of all dephosphorylation reactions, is regulated by scores of regulatory proteins, including the highly conserved SDS22 protein. SDS22 has numerous diverse functions, surprisingly acting as both a PP1 inhibitor and as an activator. Here, we integrate cellular, biophysical, and crystallographic studies to address this conundrum. We discovered that SDS22 selectively binds a unique conformation of PP1 that contains a single metal (M2) at its active site, i.e., SDS22 traps metal-deficient inactive PP1. Furthermore, we showed that SDS22 dissociation is accompanied by a second metal (M1) being loaded into PP1, as free metal cannot dissociate the complex and M1-deficient mutants remain constitutively trapped by SDS22. Together, our findings reveal that M1 metal loading and loss are essential for PP1 regulation in cells, which has broad implications for PP1 maturation, activity, and holoenzyme subunit exchange.

Authors:
 [1];  [1];  [2];  [1];  [2];  [1];  [3];  [1];  [2]; ORCiD logo [1]
  1. Univ. of Arizona, Tucson, AZ (United States)
  2. Louisiana State Univ. Health Sciences Center, Shreveport, LA (United States)
  3. Brookhaven National Lab. (BNL), Upton, NY (United States)
Publication Date:
Research Org.:
Brookhaven National Lab. (BNL), Upton, NY (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22)
OSTI Identifier:
1573470
Report Number(s):
BNL-212280-2019-JAAM
Journal ID: ISSN 0027-8424
Grant/Contract Number:  
SC0012704
Resource Type:
Accepted Manuscript
Journal Name:
Proceedings of the National Academy of Sciences of the United States of America
Additional Journal Information:
Journal Volume: 116; Journal Issue: 41; Journal ID: ISSN 0027-8424
Publisher:
National Academy of Sciences
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Choy, Meng S., Moon, Thomas M., Ravindran, Rini, Bray, Johnny A., Robinson, Lucy C., Archuleta, Tara L., Shi, Wuxian, Peti, Wolfgang, Tatchell, Kelly, and Page, Rebecca. SDS22 selectively recognizes and traps metal-deficient inactive PP1. United States: N. p., 2019. Web. doi:10.1073/pnas.1908718116.
Choy, Meng S., Moon, Thomas M., Ravindran, Rini, Bray, Johnny A., Robinson, Lucy C., Archuleta, Tara L., Shi, Wuxian, Peti, Wolfgang, Tatchell, Kelly, & Page, Rebecca. SDS22 selectively recognizes and traps metal-deficient inactive PP1. United States. doi:10.1073/pnas.1908718116.
Choy, Meng S., Moon, Thomas M., Ravindran, Rini, Bray, Johnny A., Robinson, Lucy C., Archuleta, Tara L., Shi, Wuxian, Peti, Wolfgang, Tatchell, Kelly, and Page, Rebecca. Mon . "SDS22 selectively recognizes and traps metal-deficient inactive PP1". United States. doi:10.1073/pnas.1908718116.
@article{osti_1573470,
title = {SDS22 selectively recognizes and traps metal-deficient inactive PP1},
author = {Choy, Meng S. and Moon, Thomas M. and Ravindran, Rini and Bray, Johnny A. and Robinson, Lucy C. and Archuleta, Tara L. and Shi, Wuxian and Peti, Wolfgang and Tatchell, Kelly and Page, Rebecca},
abstractNote = {The metalloenzyme protein phosphatase 1 (PP1), which is responsible for ≥50% of all dephosphorylation reactions, is regulated by scores of regulatory proteins, including the highly conserved SDS22 protein. SDS22 has numerous diverse functions, surprisingly acting as both a PP1 inhibitor and as an activator. Here, we integrate cellular, biophysical, and crystallographic studies to address this conundrum. We discovered that SDS22 selectively binds a unique conformation of PP1 that contains a single metal (M2) at its active site, i.e., SDS22 traps metal-deficient inactive PP1. Furthermore, we showed that SDS22 dissociation is accompanied by a second metal (M1) being loaded into PP1, as free metal cannot dissociate the complex and M1-deficient mutants remain constitutively trapped by SDS22. Together, our findings reveal that M1 metal loading and loss are essential for PP1 regulation in cells, which has broad implications for PP1 maturation, activity, and holoenzyme subunit exchange.},
doi = {10.1073/pnas.1908718116},
journal = {Proceedings of the National Academy of Sciences of the United States of America},
number = 41,
volume = 116,
place = {United States},
year = {2019},
month = {9}
}

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Works referenced in this record:

Protein Serine/Threonine Phosphatases: Keys to Unlocking Regulators and Substrates
journal, June 2018


Expression of Human Protein Phosphatase-1 in Saccharomyces cerevisiae Highlights the Role of Phosphatase Isoforms in Regulating Eukaryotic Functions
journal, June 2007

  • Gibbons, Jennifer A.; Kozubowski, Lukasz; Tatchell, Kelly
  • Journal of Biological Chemistry, Vol. 282, Issue 30
  • DOI: 10.1074/jbc.M701272200

Molecular Investigations of the Structure and Function of the Protein Phosphatase 1−Spinophilin−Inhibitor 2 Heterotrimeric Complex
journal, February 2011

  • Dancheck, Barbara; Ragusa, Michael J.; Allaire, Marc
  • Biochemistry, Vol. 50, Issue 7
  • DOI: 10.1021/bi101774g

Effects of stably incorporated iron on protein phosphatase‐1 structure and activity
journal, November 2018

  • Salvi, Francesca; Trebacz, Malgorzata; Kokot, Thomas
  • FEBS Letters, Vol. 592, Issue 24
  • DOI: 10.1002/1873-3468.13284

Inactivation and reactivation of phosphoprotein phosphatase
journal, January 1982

  • Yan, SauChiB.; Graves, DonaldJ.
  • Molecular and Cellular Biochemistry, Vol. 42, Issue 1
  • DOI: 10.1007/BF00223535

Activation of Protein Phosphatase 1: FORMATION OF A METALLOENZYME
journal, February 1996

  • Chu, Y.; Lee, E. Y. C.; Schlender, K. K.
  • Journal of Biological Chemistry, Vol. 271, Issue 5
  • DOI: 10.1074/jbc.271.5.2574

Three-dimensional structure of the catalytic subunit of protein serine/threonine phosphatase-1
journal, August 1995

  • Goldberg, Jonathan; Huang, Hsien-bin; Kwon, Young-guen
  • Nature, Vol. 376, Issue 6543
  • DOI: 10.1038/376745a0

Identification of the substrate recruitment mechanism of the muscle glycogen protein phosphatase 1 holoenzyme
journal, November 2018

  • Kumar, Ganesan Senthil; Choy, Meng S.; Koveal, Dorothy M.
  • Science Advances, Vol. 4, Issue 11
  • DOI: 10.1126/sciadv.aau6044

The extended PP1 toolkit: designed to create specificity
journal, August 2010

  • Bollen, Mathieu; Peti, Wolfgang; Ragusa, Michael J.
  • Trends in Biochemical Sciences, Vol. 35, Issue 8
  • DOI: 10.1016/j.tibs.2010.03.002

Regulation of protein phosphatase 1 by intrinsically disordered proteins
journal, September 2012

  • Choy, Meng S.; Page, Rebecca; Peti, Wolfgang
  • Biochemical Society Transactions, Vol. 40, Issue 5
  • DOI: 10.1042/BST20120094

Structural basis for protein phosphatase 1 regulation and specificity: Protein phosphatase 1 regulation and specificity
journal, February 2012


Understanding the antagonism of retinoblastoma protein dephosphorylation by PNUTS provides insights into the PP1 regulatory code
journal, March 2014

  • Choy, M. S.; Hieke, M.; Kumar, G. S.
  • Proceedings of the National Academy of Sciences, Vol. 111, Issue 11
  • DOI: 10.1073/pnas.1317395111

Docking Motif-Guided Mapping of the Interactome of Protein Phosphatase-1
journal, April 2009


The Molecular Mechanism of Substrate Engagement and Immunosuppressant Inhibition of Calcineurin
journal, February 2013


KNL1 Binding to PP1 and Microtubules Is Mutually Exclusive
journal, October 2018


Structural and Functional Analysis of the GADD34:PP1 eIF2α Phosphatase
journal, June 2015


Spinophilin directs protein phosphatase 1 specificity by blocking substrate binding sites
journal, March 2010

  • Ragusa, Michael J.; Dancheck, Barbara; Critton, David A.
  • Nature Structural & Molecular Biology, Vol. 17, Issue 4
  • DOI: 10.1038/nsmb.1786

Structural basis of protein phosphatase 1 regulation
journal, May 2004

  • Terrak, Mohammed; Kerff, Frederic; Langsetmo, Knut
  • Nature, Vol. 429, Issue 6993
  • DOI: 10.1038/nature02582

Systematic humanization of yeast genes reveals conserved functions and genetic modularity
journal, May 2015


Binding of the Concave Surface of the Sds22 Superhelix to the α4/α5/α6-Triangle of Protein Phosphatase-1
journal, September 2002

  • Ceulemans, Hugo; Vulsteke, Veerle; De Maeyer, Marc
  • Journal of Biological Chemistry, Vol. 277, Issue 49
  • DOI: 10.1074/jbc.M206838200

Phosphorylation of PP1 Regulator Sds22 by PLK1 Ensures Accurate Chromosome Segregation
journal, August 2016

  • Duan, Hequan; Wang, Chunli; Wang, Ming
  • Journal of Biological Chemistry, Vol. 291, Issue 40
  • DOI: 10.1074/jbc.M116.745372

Kinetochore-localized PP1–Sds22 couples chromosome segregation to polar relaxation
journal, July 2015

  • Rodrigues, Nelio T. L.; Lekomtsev, Sergey; Jananji, Silvana
  • Nature, Vol. 524, Issue 7566
  • DOI: 10.1038/nature14496

Regulators of the protein phosphatase PP1γ2, PPP1R2, PPP1R7, and PPP1R11 are involved in epididymal sperm maturation
journal, August 2018

  • Goswami, Suranjana; Korrodi‐Gregório, Luís; Sinha, Nilam
  • Journal of Cellular Physiology, Vol. 234, Issue 3
  • DOI: 10.1002/jcp.27130

Ypi1, a Positive Regulator of Nuclear Protein Phosphatase Type 1 Activity in Saccharomyces cerevisiae
journal, March 2008

  • Bharucha, Jennifer P.; Larson, Jennifer R.; Gao, Lu
  • Molecular Biology of the Cell, Vol. 19, Issue 3
  • DOI: 10.1091/mbc.E07-05-0499

The AAA-ATPase Cdc48 and cofactor Shp1 promote chromosome bi-orientation by balancing Aurora B activity
journal, May 2010

  • Cheng, Y. -L.; Chen, R. -H.
  • Journal of Cell Science, Vol. 123, Issue 12
  • DOI: 10.1242/jcs.066043

Assembly and quality control of the protein phosphatase 1 holoenzyme involves the Cdc48-Shp1 chaperone
journal, January 2015

  • Cheng, Y. -L.; Chen, R. -H.
  • Journal of Cell Science, Vol. 128, Issue 6
  • DOI: 10.1242/jcs.165159

Ubiquitin-Independent Disassembly by a p97 AAA-ATPase Complex Drives PP1 Holoenzyme Formation
journal, November 2018


The Ki-67 and RepoMan mitotic phosphatases assemble via an identical, yet novel mechanism
journal, August 2016

  • Kumar, Ganesan Senthil; Gokhan, Ezgi; De Munter, Sofie
  • eLife, Vol. 5
  • DOI: 10.7554/eLife.16539

Separation and Characterization of Two Phosphorylase Phosphatase Inhibitors from Rabbit Skeletal Muscle
journal, November 1976


Identification and Characterization of the Human HCG V Gene Product as a Novel Inhibitor of Protein Phosphatase-1
journal, November 1998

  • Zhang, Jun; Zhang, Lifang; Zhao, Sumin
  • Biochemistry, Vol. 37, Issue 47
  • DOI: 10.1021/bi981169g

Characterization of the Neuronal Targeting Protein Spinophilin and Its Interactions with Protein Phosphatase-1
journal, April 1999

  • Hsieh-Wilson, Linda C.; Allen, Patrick B.; Watanabe, Takuo
  • Biochemistry, Vol. 38, Issue 14
  • DOI: 10.1021/bi982900m

Mutational Analysis of the Catalytic Subunit of Muscle Protein Phosphatase-1
journal, January 1996

  • Zhang, Jun; Zhang, Zhongjian; Brew, Keith
  • Biochemistry, Vol. 35, Issue 20
  • DOI: 10.1021/bi952954l

The structure of SDS22 provides insights into the mechanism of heterodimer formation with PP1
journal, November 2018

  • Choy, Meng S.; Bolik-Coulon, Nicolas; Archuleta, Tara L.
  • Acta Crystallographica Section F Structural Biology Communications, Vol. 74, Issue 12
  • DOI: 10.1107/S2053230X18016503

Structure-Guided Exploration of SDS22 Interactions with Protein Phosphatase PP1 and the Splicing Factor BCLAF1
journal, March 2019


Metal ion-coordinating properties of imidazole and derivatives in aqueous solution: interrelation between complex stability and ligand basicity
journal, October 1998


Architecture, remodeling, and functions of the septin cytoskeleton
journal, August 2018

  • Marquardt, Joseph; Chen, Xi; Bi, Erfei
  • Cytoskeleton, Vol. 76, Issue 1
  • DOI: 10.1002/cm.21475

Septin structure and function in yeast and beyond
journal, March 2011


The Role of Cdc42p GTPase-activating Proteins in Assembly of the Septin Ring in Yeast
journal, October 2003

  • Caviston, Juliane P.; Longtine, Mark; Pringle, John R.
  • Molecular Biology of the Cell, Vol. 14, Issue 10
  • DOI: 10.1091/mbc.E03-04-0247

Phosphorylation-Dependent Regulation of Septin Dynamics during the Cell Cycle
journal, March 2003


Order of Stability of Metal Complexes
journal, November 1948

  • Irving, H.; Williams, R. J. P.
  • Nature, Vol. 162, Issue 4123
  • DOI: 10.1038/162746a0

Molecular modeling of the catalytic domain of serine/threonine phosphatase-1 with the Zn2+ and Mn2+ di-nuclear ion centers in the active site
journal, May 2000


Strategies to maximize heterologous protein expression in Escherichia coli with minimal cost
journal, January 2007


The Molecular Basis for Substrate Specificity of the Nuclear NIPP1:PP1 Holoenzyme
journal, October 2012


New ubiquitin-dependent mechanisms regulating the Aurora B–protein phosphatase 1 balance in Saccharomyces cerevisiae
journal, July 2018

  • Ravindran, Rini; Polk, Paula; Robinson, Lucy C.
  • Journal of Cell Science, Vol. 131, Issue 16
  • DOI: 10.1242/jcs.217620