Coordination of Substrate Binding and ATP Hydrolysis in Vps4-Mediated ESCRT-III Disassembly

Davies, Brian A.; Azmi, Ishara F.; Payne, Johanna A.; Shestakova, Anna; Horazdovsky, Bruce F.; Babst, Markus; Katzmann, David J.

doi:10.1091/mbc.e10-06-0512

Cited by 50 publications

(78 citation statements)

References 57 publications

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“…5M to P). These partially enveloped structures are exactly those expected to accumulate when neck closure and scission are inhibited by dominant negative Vps4-EQ-GFP (15,59,72,73,76,93). Interestingly, the rim or neck of the unsealed envelope is decorated by a ring of structures resembling beads on a string (Fig.…”

Section: Discussionmentioning

confidence: 53%

“…In both normal and infected cells, the final step in the ESCRT pathway is ATP hydrolysis by the AAA ATPase Vps4 at the bud neck; this drives ESCRT III dissociation and membrane scission (70)(71)(72)(73). Replacement of a glutamate by a glutamine in the Vps4 ATPase active site generates Vps4-EQ, an ATP-locked dominant negative allele (74,75) that can block ESCRT III release and arrest viral envelopment (15,59,72,76).…”

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confidence: 99%

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Herpes Simplex Virus Capsid Localization to ESCRT-VPS4 Complexes in the Presence and Absence of the Large Tegument Protein UL36p

Kharkwal

Smith

Wilson

2016

J Virol

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UL36p (VP1/2) is the largest protein encoded by herpes simplex virus 1 (HSV-1) and resides in the innermost layer of tegument, the complex protein layer between the capsid and envelope. UL36p performs multiple functions in the HSV life cycle, including a critical but unknown role in capsid cytoplasmic envelopment. We tested whether UL36p is essential for envelopment because it is required to engage capsids with the cellular ESCRT/Vps4 apparatus. A green fluorescent protein (GFP)-fused form of the dominant negative ATPase Vps4-EQ was used to irreversibly tag ESCRT envelopment sites during infection by UL36p-expressing and UL36-null HSV strains. Using fluorescence microscopy and scanning electron microscopy, we quantitated capsid/Vps4-EQ colocalization and examined the ultrastructure of the corresponding viral assembly intermediates. We found that loss of UL36p resulted in a two-thirds reduction in the efficiency of capsid/Vps4-EQ association but that the remaining UL36p-null capsids were still able to engage the ESCRT envelopment apparatus. It appears that although UL36p helps to couple HSV capsids to the ESCRT pathway, this is likely not the sole reason for its absolute requirement for envelopment. IMPORTANCEEnvelopment of the HSV capsid is essential for the assembly of an infectious virion and requires the complex interplay of a large number of viral and cellular proteins. Critical to envelope assembly is the virally encoded protein UL36p, whose function is unknown. Here we test the hypothesis that UL36p is essential for the recruitment of cellular ESCRT complexes, which are also known to be required for envelopment. Herpesviruses replicate their genomes and assemble DNApackaged capsids in the cell nucleus. It is generally accepted that capsids then bud into the inner nuclear membrane to generate primary enveloped perinuclear virions that subsequently fuse with the outer nuclear membrane, releasing mature nucleocapsids ("naked" capsids) into the cytoplasm (1-3). These capsids subsequently undergo secondary envelopment at a cytoplasmic organelle to assemble the mature, infectious virion (4-10).Herpesvirus cytoplasmic envelopment is extremely complex, requiring the coordinated interaction of multiple viral and cellular polypeptides (8,9,(11)(12)(13)(14)(15)(16)). An essential component of the envelopment apparatus is the conserved multifunctional protein UL36p (VP1/2) (17-21). UL36p is the largest structural polypeptide encoded by the members of the Herpesviridae (22, 23), and it forms the innermost layer (18, 24-32) of tegument, the complex protein scaffold between the capsid and envelope (8,16,33). UL36p attaches the capsid (18, 31, 32, 34) to multiple outer tegument components (24-30, 35, 36) that in turn bind integral membrane envelope proteins (1, 9, 37-40) and the lipid envelope (41-43). One important function of UL36p is to recruit UL37p (19,20,25,29,44,45), a putative mimic of cellular multisubunit tethering complexes (28) that mediates capsid docking to organelles, including the trans-Golgi netwo...

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Section: Discussionmentioning

confidence: 53%

mentioning

confidence: 99%

Herpes Simplex Virus Capsid Localization to ESCRT-VPS4 Complexes in the Presence and Absence of the Large Tegument Protein UL36p

Kharkwal

Smith

Wilson

2016

J Virol

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“…[30][31][32] Vps4 associates with the ESCRT-III protein complex through MIM domains, which facilitates Vps4 ATP hydrolysis and disassembly of the ESCRT-III complex during MVB formation. 9,33,34 Deletion of MIM and flanking sequences of ESCRT-III promotes complex formation and membrane association, which are visualized as enlarged vesicles/endosomes in the cytoplasm. 30,31 Chromatin modifying protein 1A (Chmp1A, yeast Did2/ Vps 46-1), which also stands for Charged multivesicular body protein 1A, is a member of the ESCRT-III complex.…”

Section: Resultsmentioning

confidence: 99%

Chromatin modifying protein 1A (Chmp1A) of the endosomal sorting complex required for transport (ESCRT)-III family activates ataxia telangiectasia mutated (ATM) for PanC-1 cell growth inhibition

Manohar

Harlow²,

Nguyen³

et al. 2011

Cell Cycle

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“…This activity of Vps4 can be tested in vitro using recombinant protein (30). Increasing concentrations of purified wild-type or Vps4 linker mutants were added to ESCRT-III-containing endosome fractions, and the resulting solubilization of the ESCRT-III subunit Snf7 was determined by centrifugation and Western blot analysis of the resulting pellet and supernatant fractions.…”

Section: Vps4 Linker Mutants Exhibit Reduced Stimulation By Escrt-iiimentioning

confidence: 99%

The Linker Region Plays a Regulatory Role in Assembly and Activity of the Vps4 AAA ATPase

Shestakova

Davies

Katzmann

et al. 2013

Journal of Biological Chemistry

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Background: Vps4 functions in eukaryotic cells in membrane fission reactions. Results: Mutational analysis suggests that the long linker region of Vps4 is not essential but regulates this ATPase. Conclusion:The binding to the substrate ESCRT-III drives formation of the active Vps4 complex. Significance: Understanding the mechanism of Vps4 function gives insights into protein trafficking, cytokinesis, and virus formation.

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Coordination of Substrate Binding and ATP Hydrolysis in Vps4-Mediated ESCRT-III Disassembly

Abstract: Vps4 disassembly of ESCRT-III plays an important role in MVB sorting, viral budding, and cytokinesis. An in vitro system was developed to investigate this process. These studies revealed new insights into the mechanisms of Vps4 function.

Cited by 50 publications

References 57 publications

Herpes Simplex Virus Capsid Localization to ESCRT-VPS4 Complexes in the Presence and Absence of the Large Tegument Protein UL36p

Herpes Simplex Virus Capsid Localization to ESCRT-VPS4 Complexes in the Presence and Absence of the Large Tegument Protein UL36p

Chromatin modifying protein 1A (Chmp1A) of the endosomal sorting complex required for transport (ESCRT)-III family activates ataxia telangiectasia mutated (ATM) for PanC-1 cell growth inhibition

The Linker Region Plays a Regulatory Role in Assembly and Activity of the Vps4 AAA ATPase

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