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Structure paper

TitleCryo-EM Structures of MDA5-dsRNA Filaments at Different Stages of ATP Hydrolysis.
Journal, issue, pagesMol Cell, Vol. 72, Issue 6, Page 999-1012.e6, Year 2018
Publish dateDec 20, 2018
AuthorsQin Yu / Kun Qu / Yorgo Modis /
PubMed AbstractDouble-stranded RNA (dsRNA) is a potent proinflammatory signature of viral infection. Long cytosolic dsRNA is recognized by MDA5. The cooperative assembly of MDA5 into helical filaments on dsRNA ...Double-stranded RNA (dsRNA) is a potent proinflammatory signature of viral infection. Long cytosolic dsRNA is recognized by MDA5. The cooperative assembly of MDA5 into helical filaments on dsRNA nucleates the assembly of a multiprotein type I interferon signaling platform. Here, we determined cryoelectron microscopy (cryo-EM) structures of MDA5-dsRNA filaments with different helical twists and bound nucleotide analogs at resolutions sufficient to build and refine atomic models. The structures identify the filament-forming interfaces, which encode the dsRNA binding cooperativity and length specificity of MDA5. The predominantly hydrophobic interface contacts confer flexibility, reflected in the variable helical twist within filaments. Mutation of filament-forming residues can result in loss or gain of signaling activity. Each MDA5 molecule spans 14 or 15 RNA base pairs, depending on the twist. Variations in twist also correlate with variations in the occupancy and type of nucleotide in the active site, providing insights on how ATP hydrolysis contributes to MDA5-dsRNA recognition.
External linksMol Cell / PubMed:30449722 / PubMed Central
MethodsEM (helical sym.)
Resolution3.68 - 4.16 Å
Structure data

0012

6gjz

EMDB-0012, PDB-6gjz:
CryoEM structure of the MDA5-dsRNA filament in complex with AMPPNP
Method: EM (helical sym.) / Resolution: 4.06 Å

0023

6gkh

EMDB-0023, PDB-6gkh:
CryoEM structure of the MDA5-dsRNA filament in complex with ADP-AlF4
Method: EM (helical sym.) / Resolution: 4.06 Å

0024

6gkm

EMDB-0024, PDB-6gkm:
CryoEM structure of the MDA5-dsRNA filament in complex with ATP (10 mM)
Method: EM (helical sym.) / Resolution: 3.87 Å

0143

6h61

EMDB-0143, PDB-6h61:
CryoEM structure of the MDA5-dsRNA filament with 89 degree twist and without nucleotide
Method: EM (helical sym.) / Resolution: 4.02 Å

0145

6h66

EMDB-0145, PDB-6h66:
CryoEM structure of the MDA5-dsRNA filament with 93 degree twist and without nucleotide
Method: EM (helical sym.) / Resolution: 4.16 Å

4338

6g19

EMDB-4338, PDB-6g19:
CryoEM structure of the MDA5-dsRNA filament with 74-degree helical twist
Method: EM (helical sym.) / Resolution: 3.68 Å

4340

6g1s

EMDB-4340, PDB-6g1s:
CryoEM structure of the MDA5-dsRNA filament with 87-degree helical twist
Method: EM (helical sym.) / Resolution: 3.93 Å

4341

6g1x

EMDB-4341, PDB-6g1x:
CryoEM structure of the MDA5-dsRNA filament with 91-degree helical twist
Method: EM (helical sym.) / Resolution: 3.93 Å

Chemicals

ChemComp-ZN:
Unknown entry

ChemComp-ANP:
PHOSPHOAMINOPHOSPHONIC ACID-ADENYLATE ESTER / AMP-PNP, energy-carrying molecule analogue*YM

ChemComp-ADP:
ADENOSINE-5'-DIPHOSPHATE / ADP, energy-carrying molecule*YM

ChemComp-ALF:
TETRAFLUOROALUMINATE ION

ChemComp-MG:
Unknown entry

ChemComp-ATP:
ADENOSINE-5'-TRIPHOSPHATE / ATP, energy-carrying molecule*YM

Source
  • mus musculus (house mouse)
  • Pseudomonas savastanoi pv. phaseolicola (bacteria)
  • pseudomonas phage phi6 (bacteriophage)
KeywordsIMMUNE SYSTEM / Protein-RNA complex / helical filament / ATPase / innate immune receptor

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