A Crescent-shaped ALIX Dimer Targets ESCRT-III CHMP4 Filaments

Overview

Journal Structure

Publisher Cell Press

Specialties Biochemistry
Biotechnology
Cell Biology
Molecular Biology

Date 2009 Jun 16

PMID 19523902

Citations 81

Authors

Ricardo Pires

Bettina Hartlieb

Luca Signor

Guy Schoehn

Suman Lata

Manfred Roessle

Christine Moriscot

Sergei Popov

Andreas Hinz

Marc Jamin

Veronique Boyer

Remy Sadoul

Eric Forest

Dmitri I Svergun

Heinrich G Gottlinger

Winfried Weissenhorn

Affiliations

Soon will be listed here.

Abstract

ALIX recruits ESCRT-III CHMP4 and is involved in membrane remodeling during endosomal receptor sorting, budding of some enveloped viruses, and cytokinesis. We show that ALIX dimerizes via the middle domain (ALIX(-V)) in solution. Structural modeling based on small angle X-ray scattering (SAXS) data reveals an elongated crescent-shaped conformation for dimeric ALIX lacking the proline-rich domain (ALIX(BRO1-V)). Mutations at the dimerization interface prevent dimerization and induce an open elongated monomeric conformation of ALIX(-V) as determined by SAXS modeling. ALIX dimerizes in vivo and dimeric ALIX colocalizes with CHMP4B upon coexpression. We show further that ALIX dimerization affects HIV-1 budding. C-terminally truncated activated CHMP4B retaining the ALIX binding site forms linear, circular, and helical filaments in vitro, which can be bridged by ALIX. Our data suggest that dimeric ALIX represents the active form that interacts with ESCRT-III CHMP4 polymers and functions as a scaffolding protein during membrane remodeling processes.

Citing Articles

NEDD4 family ubiquitin ligase AIP4 interacts with Alix to enable HBV naked capsid egress in an Alix ubiquitination-independent manner.

Shen S, Cai D, Liang H, Zeng G, Liu W, Yan R PLoS Pathog. 2024; 20(9):e1012485.

PMID: 39259704 PMC: 11389946. DOI: 10.1371/journal.ppat.1012485.

: EhADH, an Alix Protein, Participates in Several Virulence Events through Its Different Domains.

Zanatta D, Betanzos A, Azuara-Liceaga E, Montano S, Orozco E Int J Mol Sci. 2024; 25(14).

PMID: 39062867 PMC: 11277477. DOI: 10.3390/ijms25147609.

Palmitoylation of vacuole membrane protein 1 promotes small extracellular vesicle secretion via interaction with ALIX and influences intercellular communication.

Qu M, Liu X, Wang X, Li Z, Zhou L, Li H Cell Commun Signal. 2024; 22(1):150.

PMID: 38403678 PMC: 10895845. DOI: 10.1186/s12964-024-01529-6.

An Inducible ESCRT-III Inhibition Tool to Control HIV-1 Budding.

Wang H, Gallet B, Moriscot C, Pezet M, Chatellard C, Kleman J Viruses. 2023; 15(12).

PMID: 38140530 PMC: 10748027. DOI: 10.3390/v15122289.

SUMOylation-triggered ALIX activation modulates extracellular vesicles circTLCD4-RWDD3 to promote lymphatic metastasis of non-small cell lung cancer.

Diao X, Guo C, Zheng H, Zhao K, Luo Y, An M Signal Transduct Target Ther. 2023; 8(1):426.

PMID: 37925421 PMC: 10625632. DOI: 10.1038/s41392-023-01685-0.

References

Vito P, Pellegrini L, Guiet C, DAdamio L . Cloning of AIP1, a novel protein that associates with the apoptosis-linked gene ALG-2 in a Ca2+-dependent reaction. J Biol Chem. 1999; 274(3):1533-40. DOI: 10.1074/jbc.274.3.1533. View

Garrus J, von Schwedler U, Pornillos O, Morham S, Zavitz K, Wang H . Tsg101 and the vacuolar protein sorting pathway are essential for HIV-1 budding. Cell. 2001; 107(1):55-65. DOI: 10.1016/s0092-8674(01)00506-2. View

Gaillard J, Neumann E, Van Damme D, Stoppin-Mellet V, Ebel C, Barbier E . Two microtubule-associated proteins of Arabidopsis MAP65s promote antiparallel microtubule bundling. Mol Biol Cell. 2008; 19(10):4534-44. PMC: 2555953. DOI: 10.1091/mbc.e08-04-0341. View

Gruenberg J, Stenmark H . The biogenesis of multivesicular endosomes. Nat Rev Mol Cell Biol. 2004; 5(4):317-23. DOI: 10.1038/nrm1360. View

Fisher R, Chung H, Zhai Q, Robinson H, Sundquist W, Hill C . Structural and biochemical studies of ALIX/AIP1 and its role in retrovirus budding. Cell. 2007; 128(5):841-52. DOI: 10.1016/j.cell.2007.01.035. View

Lin Y, Kimpler L, Naismith T, Lauer J, Hanson P . Interaction of the mammalian endosomal sorting complex required for transport (ESCRT) III protein hSnf7-1 with itself, membranes, and the AAA+ ATPase SKD1. J Biol Chem. 2005; 280(13):12799-809. DOI: 10.1074/jbc.M413968200. View

Zhou X, Pan S, Sun L, Corvera J, Lin S, Kuang J . The HIV-1 p6/EIAV p9 docking site in Alix is autoinhibited as revealed by a conformation-sensitive anti-Alix monoclonal antibody. Biochem J. 2008; 414(2):215-20. DOI: 10.1042/BJ20080642. View

Chatellard-Causse C, Blot B, Cristina N, Torch S, Missotten M, Sadoul R . Alix (ALG-2-interacting protein X), a protein involved in apoptosis, binds to endophilins and induces cytoplasmic vacuolization. J Biol Chem. 2002; 277(32):29108-15. DOI: 10.1074/jbc.M204019200. View

Tanzi G, Piefer A, Bates P . Equine infectious anemia virus utilizes host vesicular protein sorting machinery during particle release. J Virol. 2003; 77(15):8440-7. PMC: 165230. DOI: 10.1128/jvi.77.15.8440-8447.2003. View

10.

Missotten M, Nichols A, Rieger K, Sadoul R . Alix, a novel mouse protein undergoing calcium-dependent interaction with the apoptosis-linked-gene 2 (ALG-2) protein. Cell Death Differ. 1999; 6(2):124-9. DOI: 10.1038/sj.cdd.4400456. View

11.

Zhai Q, Fisher R, Chung H, Myszka D, Sundquist W, Hill C . Structural and functional studies of ALIX interactions with YPX(n)L late domains of HIV-1 and EIAV. Nat Struct Mol Biol. 2007; 15(1):43-9. DOI: 10.1038/nsmb1319. View

12.

Martin-Serrano J, Yarovoy A, Perez-Caballero D, Bieniasz P, Yaravoy A . Divergent retroviral late-budding domains recruit vacuolar protein sorting factors by using alternative adaptor proteins. Proc Natl Acad Sci U S A. 2003; 100(21):12414-9. PMC: 218772. DOI: 10.1073/pnas.2133846100. View

13.

Cabezas A, Bache K, Brech A, Stenmark H . Alix regulates cortical actin and the spatial distribution of endosomes. J Cell Sci. 2005; 118(Pt 12):2625-35. DOI: 10.1242/jcs.02382. View

14.

Hicke L . Protein regulation by monoubiquitin. Nat Rev Mol Cell Biol. 2001; 2(3):195-201. DOI: 10.1038/35056583. View

15.

Zhou X, Pan S, Sun L, Corvera J, Lee Y, Lin S . The CHMP4b- and Src-docking sites in the Bro1 domain are autoinhibited in the native state of Alix. Biochem J. 2008; 418(2):277-84. DOI: 10.1042/BJ20081388. View

16.

McCullough J, Fisher R, Whitby F, Sundquist W, Hill C . ALIX-CHMP4 interactions in the human ESCRT pathway. Proc Natl Acad Sci U S A. 2008; 105(22):7687-91. PMC: 2409388. DOI: 10.1073/pnas.0801567105. View

17.

Babst M, Katzmann D, Meerloo T, Emr S . Escrt-III: an endosome-associated heterooligomeric protein complex required for mvb sorting. Dev Cell. 2002; 3(2):271-82. DOI: 10.1016/s1534-5807(02)00220-4. View

18.

Weissenhorn W . Crystal structure of the endophilin-A1 BAR domain. J Mol Biol. 2005; 351(3):653-61. DOI: 10.1016/j.jmb.2005.06.013. View

19.

Hanson P, Roth R, Lin Y, Heuser J . Plasma membrane deformation by circular arrays of ESCRT-III protein filaments. J Cell Biol. 2008; 180(2):389-402. PMC: 2213594. DOI: 10.1083/jcb.200707031. View

20.

Morita E, Sandrin V, Chung H, Morham S, Gygi S, Rodesch C . Human ESCRT and ALIX proteins interact with proteins of the midbody and function in cytokinesis. EMBO J. 2007; 26(19):4215-27. PMC: 2230844. DOI: 10.1038/sj.emboj.7601850. View