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Title: BECN2 interacts with ATG14 through a metastable coiled-coil to mediate autophagy: BECN2 CCD Structure and Interaction with ATG14

Abstract

ATG14 binding to BECN/Beclin homologs is essential for autophagy, a critical catabolic homeostasis pathway. Here, we show that the α-helical, coiled-coil domain (CCD) of BECN2, a recently identified mammalian BECN1 paralog, forms an antiparallel, curved homodimer with seven pairs of nonideal packing interactions, while the BECN2 CCD and ATG14 CCD form a parallel, curved heterodimer stabilized by multiple, conserved polar interactions. Compared to BECN1, the BECN2 CCD forms a weaker homodimer, but binds more tightly to the ATG14 CCD. Mutation of nonideal BECN2 interface residues to more ideal pairs improves homodimer self-association and thermal stability. Unlike BECN1, all BECN2 CCD mutants bind ATG14, although more weakly than wild type. Thus, polar BECN2 CCD interface residues result in a metastable homodimer, facilitating dissociation, but enable better interactions with polar ATG14 residues stabilizing the BECN2:ATG14 heterodimer. These structure-based mechanistic differences in BECN1 and BECN2 homodimerization and heterodimerization likely dictate competitive ATG14 recruitment.

Authors:
 [1];  [1];  [1];  [2];  [3];  [4];  [1];  [1]
  1. Department of Chemistry and Biochemistry, North Dakota State University, Fargo North Dakota 58108-6050
  2. Department of Chemistry and Chemical Biology, Cornell University, Northeastern Collaborative Access Team, Argonne National Laboratory, Argonne Illinois 60439
  3. Biophysics Collaborative Access Team/Illinois Institute of Technology, Advanced Photon Source, Argonne National Laboratory, Argonne Illinois 60439
  4. Center for Autophagy Research, Department of Internal Medicine and Howard Hughes Medical Research Institute, University of Texas Southwestern Medical Center, Dallas Texas 75390
Publication Date:
Research Org.:
Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
Sponsoring Org.:
National Institutes of Health (NIH)
OSTI Identifier:
1418032
Resource Type:
Journal Article
Journal Name:
Protein Science
Additional Journal Information:
Journal Volume: 26; Journal Issue: 5; Journal ID: ISSN 0961-8368
Publisher:
The Protein Society
Country of Publication:
United States
Language:
ENGLISH
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Su, Minfei, Li, Yue, Wyborny, Shane, Neau, David, Chakravarthy, Srinivas, Levine, Beth, Colbert, Christopher L., and Sinha, Sangita C. BECN2 interacts with ATG14 through a metastable coiled-coil to mediate autophagy: BECN2 CCD Structure and Interaction with ATG14. United States: N. p., 2017. Web. doi:10.1002/pro.3140.
Su, Minfei, Li, Yue, Wyborny, Shane, Neau, David, Chakravarthy, Srinivas, Levine, Beth, Colbert, Christopher L., & Sinha, Sangita C. BECN2 interacts with ATG14 through a metastable coiled-coil to mediate autophagy: BECN2 CCD Structure and Interaction with ATG14. United States. doi:10.1002/pro.3140.
Su, Minfei, Li, Yue, Wyborny, Shane, Neau, David, Chakravarthy, Srinivas, Levine, Beth, Colbert, Christopher L., and Sinha, Sangita C. Sun . "BECN2 interacts with ATG14 through a metastable coiled-coil to mediate autophagy: BECN2 CCD Structure and Interaction with ATG14". United States. doi:10.1002/pro.3140.
@article{osti_1418032,
title = {BECN2 interacts with ATG14 through a metastable coiled-coil to mediate autophagy: BECN2 CCD Structure and Interaction with ATG14},
author = {Su, Minfei and Li, Yue and Wyborny, Shane and Neau, David and Chakravarthy, Srinivas and Levine, Beth and Colbert, Christopher L. and Sinha, Sangita C.},
abstractNote = {ATG14 binding to BECN/Beclin homologs is essential for autophagy, a critical catabolic homeostasis pathway. Here, we show that the α-helical, coiled-coil domain (CCD) of BECN2, a recently identified mammalian BECN1 paralog, forms an antiparallel, curved homodimer with seven pairs of nonideal packing interactions, while the BECN2 CCD and ATG14 CCD form a parallel, curved heterodimer stabilized by multiple, conserved polar interactions. Compared to BECN1, the BECN2 CCD forms a weaker homodimer, but binds more tightly to the ATG14 CCD. Mutation of nonideal BECN2 interface residues to more ideal pairs improves homodimer self-association and thermal stability. Unlike BECN1, all BECN2 CCD mutants bind ATG14, although more weakly than wild type. Thus, polar BECN2 CCD interface residues result in a metastable homodimer, facilitating dissociation, but enable better interactions with polar ATG14 residues stabilizing the BECN2:ATG14 heterodimer. These structure-based mechanistic differences in BECN1 and BECN2 homodimerization and heterodimerization likely dictate competitive ATG14 recruitment.},
doi = {10.1002/pro.3140},
journal = {Protein Science},
issn = {0961-8368},
number = 5,
volume = 26,
place = {United States},
year = {2017},
month = {3}
}

Works referenced in this record:

The BECN1 N-terminal domain is intrinsically disordered
journal, March 2016


Crystal structure and biochemical analyses reveal Beclin 1 as a novel membrane binding protein
journal, February 2012

  • Huang, Weijiao; Choi, Wooyoung; Hu, Wanqiu
  • Cell Research, Vol. 22, Issue 3
  • DOI: 10.1038/cr.2012.24

Bcl-x L and UVRAG Cause a Monomer-Dimer Switch in Beclin1
journal, July 2008

  • Noble, Christian G.; Dong, Jing-Ming; Manser, Edward
  • Journal of Biological Chemistry, Vol. 283, Issue 38
  • DOI: 10.1074/jbc.M804723200

Electrostatics of nanosystems: Application to microtubules and the ribosome
journal, August 2001

  • Baker, N. A.; Sept, D.; Joseph, S.
  • Proceedings of the National Academy of Sciences, Vol. 98, Issue 18, p. 10037-10041
  • DOI: 10.1073/pnas.181342398

Autophagy and Its Possible Roles in Nervous System Diseases, Damage and Repair
journal, April 2005

  • Rubinsztein, David C.; DiFiglia, Marian; Heintz, Nathaniel
  • Autophagy, Vol. 1, Issue 1
  • DOI: 10.4161/auto.1.1.1513

Two Beclin 1-binding proteins, Atg14L and Rubicon, reciprocally regulate autophagy at different stages
journal, March 2009

  • Matsunaga, Kohichi; Saitoh, Tatsuya; Tabata, Keisuke
  • Nature Cell Biology, Vol. 11, Issue 4
  • DOI: 10.1038/ncb1846

Autophagy in the Pathogenesis of Disease
journal, January 2008


Autophagy in mammalian development and differentiation
journal, September 2010

  • Mizushima, Noboru; Levine, Beth
  • Nature Cell Biology, Vol. 12, Issue 9
  • DOI: 10.1038/ncb0910-823

Intrinsically disordered regions in autophagy proteins: Intrinsically Disordered Protein in Autophagy
journal, October 2013

  • Mei, Yang; Su, Minfei; Soni, Gaurav
  • Proteins: Structure, Function, and Bioinformatics, Vol. 82, Issue 4
  • DOI: 10.1002/prot.24424

Development by Self-Digestion
journal, April 2004


Global Rigid Body Modeling of Macromolecular Complexes against Small-Angle Scattering Data
journal, August 2005


Coiled Coil Domains: Stability, Specificity, and Biological Implications
journal, January 2004


Targeting γ-Herpesvirus 68 Bcl-2-mediated Down-regulation of Autophagy
journal, January 2014

  • Su, Minfei; Mei, Yang; Sanishvili, Ruslan
  • Journal of Biological Chemistry, Vol. 289, Issue 12
  • DOI: 10.1074/jbc.M113.515361

The tripartite motif coiled-coil is an elongated antiparallel hairpin dimer
journal, February 2014

  • Sanchez, J. G.; Okreglicka, K.; Chandrasekaran, V.
  • Proceedings of the National Academy of Sciences, Vol. 111, Issue 7
  • DOI: 10.1073/pnas.1318962111

Structure of the Novel C-terminal Domain of Vacuolar Protein Sorting 30/Autophagy-related Protein 6 and Its Specific Role in Autophagy
journal, March 2012

  • Noda, Nobuo N.; Kobayashi, Takafumi; Adachi, Wakana
  • Journal of Biological Chemistry, Vol. 287, Issue 20
  • DOI: 10.1074/jbc.M112.348250

Architecture and dynamics of the autophagic phosphatidylinositol 3-kinase complex
journal, December 2014

  • Baskaran, Sulochanadevi; Carlson, Lars-Anders; Stjepanovic, Goran
  • eLife, Vol. 3
  • DOI: 10.7554/eLife.05115

Autophagy and Aging
journal, September 2011


Structure and flexibility of the endosomal Vps34 complex reveals the basis of its function on membranes
journal, October 2015


Conformational flexibility of BECN1: Essential to its key role in autophagy and beyond: BECN1 Structure and Mechanism
journal, August 2016

  • Mei, Yang; Glover, Karen; Su, Minfei
  • Protein Science, Vol. 25, Issue 10
  • DOI: 10.1002/pro.2984

Identification of BECN1 and ATG14 Coiled-Coil Interface Residues That Are Important for Starvation-Induced Autophagy
journal, July 2016


Beclin 2 Functions in Autophagy, Degradation of G Protein-Coupled Receptors, and Metabolism
journal, August 2013


Beclin 1 self-association is independent of autophagy induction by amino acid deprivation and rapamycin treatment
journal, May 2010

  • Adi-Harel, Shelly; Erlich, Shlomit; Schmukler, Eran
  • Journal of Cellular Biochemistry, Vol. 110, Issue 5
  • DOI: 10.1002/jcb.22642

Conformational Flexibility Enables the Function of a BECN1 Region Essential for Starvation-Mediated Autophagy
journal, March 2016


CCBuilder: an interactive web-based tool for building, designing and assessing coiled-coil protein assemblies
journal, July 2014


(PS) 2 : protein structure prediction server version 3.0
journal, May 2015

  • Huang, Tsun-Tsao; Hwang, Jenn-Kang; Chen, Chu-Huang
  • Nucleic Acids Research, Vol. 43, Issue W1
  • DOI: 10.1093/nar/gkv454

Imperfect interface of Beclin1 coiled-coil domain regulates homodimer and heterodimer formation with Atg14L and UVRAG
journal, January 2012

  • Li, Xiaohua; He, Liqiang; Che, Ka Hing
  • Nature Communications, Vol. 3, Issue 1
  • DOI: 10.1038/ncomms1648

Crystal Structure of the Bcl-X L -Beclin 1 Peptide Complex : BECLIN 1 IS A NOVEL BH3-ONLY PROTEIN
journal, March 2007

  • Oberstein, Adam; Jeffrey, Philip D.; Shi, Yigong
  • Journal of Biological Chemistry, Vol. 282, Issue 17
  • DOI: 10.1074/jbc.M700492200

Autophagy: process and function
journal, November 2007


The autophagy effector Beclin 1: a novel BH3-only protein
journal, December 2008


Structural and Biochemical Bases for the Inhibition of Autophagy and Apoptosis by Viral BCL-2 of Murine γ-Herpesvirus 68
journal, January 2008


Autophagy: molecular machinery for self-eating
journal, October 2005


Molecular Basis of Bcl-xL’s Target Recognition Versatility Revealed by the Structure of Bcl-xL in Complex with the BH3 Domain of Beclin-1
journal, September 2007


Molecular basis of the regulation of Beclin 1-dependent autophagy by the γ-herpesvirus 68 Bcl-2 homolog M11
journal, November 2008

  • Sinha, Sangita C.; Colbert, Christopher L.; Becker, Nils
  • Autophagy, Vol. 4, Issue 8
  • DOI: 10.4161/auto.6803

Beclin 1 Forms Two Distinct Phosphatidylinositol 3-Kinase Complexes with Mammalian Atg14 and UVRAG
journal, December 2008

  • Itakura, Eisuke; Kishi, Chieko; Inoue, Kinji
  • Molecular Biology of the Cell, Vol. 19, Issue 12
  • DOI: 10.1091/mbc.E08-01-0080

Endolysosomal trafficking of viral G protein-coupled receptor functions in innate immunity and control of viral oncogenesis
journal, February 2016

  • Dong, Xiaonan; Cheng, Adam; Zou, Zhongju
  • Proceedings of the National Academy of Sciences, Vol. 113, Issue 11
  • DOI: 10.1073/pnas.1601860113

Autophagy in immunity and inflammation
journal, January 2011

  • Levine, Beth; Mizushima, Noboru; Virgin, Herbert W.
  • Nature, Vol. 469, Issue 7330
  • DOI: 10.1038/nature09782

Identification of Barkor as a mammalian autophagy-specific factor for Beclin 1 and class III phosphatidylinositol 3-kinase
journal, December 2008

  • Sun, Q.; Fan, W.; Chen, K.
  • Proceedings of the National Academy of Sciences, Vol. 105, Issue 49
  • DOI: 10.1073/pnas.0810452105

Bcl-2 Antiapoptotic Proteins Inhibit Beclin 1-Dependent Autophagy
journal, September 2005