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Structure paper

TitleMultiple conformations facilitate PilT function in the type IV pilus.
Journal, issue, pagesNat Commun, Vol. 10, Issue 1, Page 5198, Year 2019
Publish dateNov 15, 2019
AuthorsMatthew McCallum / Samir Benlekbir / Sheryl Nguyen / Stephanie Tammam / John L Rubinstein / Lori L Burrows / P Lynne Howell /
PubMed AbstractType IV pilus-like systems are protein complexes that polymerize pilin fibres. They are critical for virulence in many bacterial pathogens. Pilin polymerization and depolymerization are powered by ...Type IV pilus-like systems are protein complexes that polymerize pilin fibres. They are critical for virulence in many bacterial pathogens. Pilin polymerization and depolymerization are powered by motor ATPases of the PilT/VirB11-like family. This family is thought to operate with C symmetry; however, most of these ATPases crystallize with either C or C symmetric conformations. The relevance of these conformations is unclear. Here, we determine the X-ray structures of PilT in four unique conformations and use these structures to classify the conformation of available PilT/VirB11-like family member structures. Single particle electron cryomicroscopy (cryoEM) structures of PilT reveal condition-dependent preferences for C C, and C conformations. The physiologic importance of these conformations is validated by coevolution analysis and functional studies of point mutants, identifying a rare gain-of-function mutation that favours the C conformation. With these data, we propose a comprehensive model of PilT function with broad implications for PilT/VirB11-like family members.
External linksNat Commun / PubMed:31729381 / PubMed Central
MethodsEM (single particle) / X-ray diffraction
Resolution1.892 - 7.8 Å
Structure data

20114

6olj

EMDB-20114, PDB-6olj:
CryoEM structure of PilB from Geobacter metallireducens: C2ccocco conformation
Method: EM (single particle) / Resolution: 7.8 Å

20115

6olk

EMDB-20115, PDB-6olk:
CryoEM structure of PilT4 from Geobacter metallireducens without adding nucleotide: C3ocococ conformation
Method: EM (single particle) / Resolution: 4.0 Å

20116

6oll

EMDB-20116, PDB-6oll:
CryoEM structure of PilT4 from Geobacter metallireducens without adding nucleotide: C2oocooc conformation
Method: EM (single particle) / Resolution: 4.1 Å

20117

6olm

EMDB-20117, PDB-6olm:
CryoEM structure of PilT4 from Geobacter metallireducens with added ATP: C6cccccc conformation
Method: EM (single particle) / Resolution: 4.4 Å

PDB-6ojx:
PilT4 from Geobacter metallireducens bound to ATP
Method: X-RAY DIFFRACTION / Resolution: 1.892 Å

PDB-6ojy:
Methylated PilT4 from Geobacter metallireducens bound to sulfate: C3ocococ conformation
Method: X-RAY DIFFRACTION / Resolution: 3.3 Å

PDB-6ojz:
PilT4 from Geobacter metallireducens bound to ADP with partial occupancy: C3ocococ conformation
Method: X-RAY DIFFRACTION / Resolution: 3.027 Å

PDB-6ok2:
PilT4 from Geobacter metallireducens bound to ADP: C3ocococ conformation
Method: X-RAY DIFFRACTION / Resolution: 3.287 Å

PDB-6okv:
PilT4 from Geobacter metallireducens bound to AMP-PNP: C2ccocco conformation
Method: X-RAY DIFFRACTION / Resolution: 4.007 Å

Chemicals

ChemComp-EDO:
1,2-ETHANEDIOL / Ethylene glycol

ChemComp-MG:
Unknown entry

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

ChemComp-HOH:
WATER / Water

ChemComp-SO4:
SULFATE ION / Sulfate

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

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

Source
  • Geobacter metallireducens (bacteria)
  • geobacter metallireducens (strain gs-15 / atcc 53774 / dsm 7210) (bacteria)
KeywordsMOTOR PROTEIN / T4P / ATPase / type IV pilus / motor

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