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- EMDB-25866: L. monocytogenes GS(14)-Q-GlnR peptide -

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Basic information

Entry
Database: EMDB / ID: EMD-25866
TitleL. monocytogenes GS(14)-Q-GlnR peptide
Map dataSharpened (B factor 110.5)
Sample
  • Complex: Tetradecameric L. monocytogenes GS complex with glutamine and GlnR C-tail peptides
    • Protein or peptide: Glutamine synthetase
    • Protein or peptide: C-tail peptide of Glutamine synthetase repressor
  • Ligand: MAGNESIUM ION
  • Ligand: GLUTAMINE
Keywordsglutamine synthetase repressor tetradecamer / BIOSYNTHETIC PROTEIN / LIGASE
Function / homology
Function and homology information


glutamine synthetase / glutamine biosynthetic process / glutamine synthetase activity / regulation of DNA-templated transcription / DNA binding / ATP binding / metal ion binding / cytoplasm
Similarity search - Function
MerR-type HTH domain signature. / Glutamine synthetase type I / Glutamine synthetase (GS) beta-grasp domain profile. / Glutamine synthetase (GS) catalytic domain profile. / Glutamine synthetase, N-terminal conserved site / Glutamine synthetase signature 1. / Glutamine synthetase, beta-Grasp domain / MerR HTH family regulatory protein / MerR-type HTH domain profile. / helix_turn_helix, mercury resistance ...MerR-type HTH domain signature. / Glutamine synthetase type I / Glutamine synthetase (GS) beta-grasp domain profile. / Glutamine synthetase (GS) catalytic domain profile. / Glutamine synthetase, N-terminal conserved site / Glutamine synthetase signature 1. / Glutamine synthetase, beta-Grasp domain / MerR HTH family regulatory protein / MerR-type HTH domain profile. / helix_turn_helix, mercury resistance / Glutamine synthetase, glycine-rich site / Glutamine synthetase putative ATP-binding region signature. / MerR-type HTH domain / Glutamine synthetase, N-terminal domain / Glutamine synthetase, N-terminal domain superfamily / Glutamine synthetase, catalytic domain / Glutamine synthetase, catalytic domain / Glutamine synthetase, catalytic domain / Glutamine synthetase/guanido kinase, catalytic domain / Putative DNA-binding domain superfamily
Similarity search - Domain/homology
Glutamine synthetase / Glutamine synthetase repressor
Similarity search - Component
Biological speciesListeria monocytogenes (bacteria)
Methodsingle particle reconstruction / cryo EM / Resolution: 2.61 Å
AuthorsTravis BA / Peck J
Funding support United States, 3 items
OrganizationGrant numberCountry
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)R35-GM130290 United States
National Institutes of Health/National Institute Of Allergy and Infectious Diseases (NIH/NIAID)F31-AI150138 United States
National Institutes of Health/National Institute of Environmental Health Sciences (NIH/NIEHS)ZIC-ES103326 United States
CitationJournal: Nat Commun / Year: 2022
Title: Molecular dissection of the glutamine synthetase-GlnR nitrogen regulatory circuitry in Gram-positive bacteria.
Authors: Brady A Travis / Jared V Peck / Raul Salinas / Brandon Dopkins / Nicholas Lent / Viet D Nguyen / Mario J Borgnia / Richard G Brennan / Maria A Schumacher /
Abstract: How bacteria sense and respond to nitrogen levels are central questions in microbial physiology. In Gram-positive bacteria, nitrogen homeostasis is controlled by an operon encoding glutamine ...How bacteria sense and respond to nitrogen levels are central questions in microbial physiology. In Gram-positive bacteria, nitrogen homeostasis is controlled by an operon encoding glutamine synthetase (GS), a dodecameric machine that assimilates ammonium into glutamine, and the GlnR repressor. GlnR detects nitrogen excess indirectly by binding glutamine-feedback-inhibited-GS (FBI-GS), which activates its transcription-repression function. The molecular mechanisms behind this regulatory circuitry, however, are unknown. Here we describe biochemical and structural analyses of GS and FBI-GS-GlnR complexes from pathogenic and non-pathogenic Gram-positive bacteria. The structures show FBI-GS binds the GlnR C-terminal domain within its active-site cavity, juxtaposing two GlnR monomers to form a DNA-binding-competent GlnR dimer. The FBI-GS-GlnR interaction stabilizes the inactive GS conformation. Strikingly, this interaction also favors a remarkable dodecamer to tetradecamer transition in some GS, breaking the paradigm that all bacterial GS are dodecamers. These data thus unveil unique structural mechanisms of transcription and enzymatic regulation.
History
DepositionJan 6, 2022-
Header (metadata) releaseJun 29, 2022-
Map releaseJun 29, 2022-
UpdateFeb 28, 2024-
Current statusFeb 28, 2024Processing site: RCSB / Status: Released

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Structure visualization

Supplemental images

emd_25866.png

Downloads & links

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Map

FileDownload / File: emd_25866.map.gz / Format: CCP4 / Size: 244.1 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationSharpened (B factor 110.5)
Voxel sizeX=Y=Z: 0.88 Å
Density
Contour LevelBy AUTHOR: 0.5
Minimum - Maximum-1.6861023 - 2.25136
Average (Standard dev.)0.0007041595 (±0.0729491)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions400400400
Spacing400400400
CellA=B=C: 352.0 Å
α=β=γ: 90.0 °

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Supplemental data

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Additional map: unsharpened

Fileemd_25866_additional_1.map
Annotationunsharpened
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Sample components

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Entire : Tetradecameric L. monocytogenes GS complex with glutamine and Gln...

EntireName: Tetradecameric L. monocytogenes GS complex with glutamine and GlnR C-tail peptides
Components
  • Complex: Tetradecameric L. monocytogenes GS complex with glutamine and GlnR C-tail peptides
    • Protein or peptide: Glutamine synthetase
    • Protein or peptide: C-tail peptide of Glutamine synthetase repressor
  • Ligand: MAGNESIUM ION
  • Ligand: GLUTAMINE

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Supramolecule #1: Tetradecameric L. monocytogenes GS complex with glutamine and Gln...

SupramoleculeName: Tetradecameric L. monocytogenes GS complex with glutamine and GlnR C-tail peptides
type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#2

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Macromolecule #1: Glutamine synthetase

MacromoleculeName: Glutamine synthetase / type: protein_or_peptide / ID: 1 / Number of copies: 14 / Enantiomer: LEVO / EC number: glutamine synthetase
Source (natural)Organism: Listeria monocytogenes (bacteria)
Molecular weightTheoretical: 52.763766 KDa
Recombinant expressionOrganism: Escherichia coli (E. coli)
SequenceString: MGSSHHHHHH SSGLVPRGSH MAKYTKEDIF RFADEQNVKF IRLQFTDILG IIKNVEIPVS QLKKALDNKI MFDGSSIEGF VRIEESDMY LFPDLDTWVV FPWTAEKGKV ARMICDIYNP DMTPFAGDPR ANLKRVLKEM EELGFTEFNL GPEPEFFLFK L DENRRPTL ...String:
MGSSHHHHHH SSGLVPRGSH MAKYTKEDIF RFADEQNVKF IRLQFTDILG IIKNVEIPVS QLKKALDNKI MFDGSSIEGF VRIEESDMY LFPDLDTWVV FPWTAEKGKV ARMICDIYNP DMTPFAGDPR ANLKRVLKEM EELGFTEFNL GPEPEFFLFK L DENRRPTL ELNDSGGYFD LAPTDLGENC RRDIVLELEE MGFEIEASHH EVAPGQHEID FKYEDAITAC DSIQTFKLVV KT IARKHGL HATFMPKPLF GVNGSGMHFN MSLFNEKGNA FFDESGELEL SQTAYHFLAG MLKHARGYTA VTNPTINSFK RLV PGYEAP CYIAWSGKNR SPLVRVPSSR GLSTRLELRS VDPSANPYLA MAVLLKAGLS GIKDELTPPA PVDRNIYGMN EEER EATGI YDLPESLGHA LIELEKNEII KDGLGEHIFE HFIEAKTIEC DMFRTAVHPW EREQYLEIY

UniProtKB: Glutamine synthetase

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Macromolecule #2: C-tail peptide of Glutamine synthetase repressor

MacromoleculeName: C-tail peptide of Glutamine synthetase repressor / type: protein_or_peptide / ID: 2 / Number of copies: 14 / Enantiomer: LEVO
Source (natural)Organism: Listeria monocytogenes (bacteria)
Molecular weightTheoretical: 660.784 Da
SequenceString:
QLPRF

UniProtKB: Glutamine synthetase repressor

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Macromolecule #3: MAGNESIUM ION

MacromoleculeName: MAGNESIUM ION / type: ligand / ID: 3 / Number of copies: 28 / Formula: MG
Molecular weightTheoretical: 24.305 Da

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Macromolecule #4: GLUTAMINE

MacromoleculeName: GLUTAMINE / type: ligand / ID: 4 / Number of copies: 14 / Formula: GLN
Molecular weightTheoretical: 146.144 Da
Chemical component information

ChemComp-GLN:
GLUTAMINE / Glutamine

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Experimental details

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Structure determination

Methodcryo EM
Processingsingle particle reconstruction
Aggregation stateparticle

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Sample preparation

Concentration0.9 mg/mL
BufferpH: 7.5
VitrificationCryogen name: ETHANE

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Electron microscopy

MicroscopeFEI TALOS ARCTICA
Electron beamAcceleration voltage: 200 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELDBright-field microscopy / Nominal defocus max: 2.2 µm / Nominal defocus min: 0.3 µm
Image recordingFilm or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Average electron dose: 42.65 e/Å2
Experimental equipment
Model: Talos Arctica / Image courtesy: FEI Company

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Image processing

Startup modelType of model: NONE
Initial angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: cryoSPARC (ver. 3.2)
Final angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: cryoSPARC (ver. 3.2)
Final reconstructionApplied symmetry - Point group: D7 (2x7 fold dihedral) / Resolution.type: BY AUTHOR / Resolution: 2.61 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: cryoSPARC (ver. 3.2) / Number images used: 212272

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Atomic model buiding 1

RefinementSpace: REAL
Output model

PDB-7tf9:
L. monocytogenes GS(14)-Q-GlnR peptide

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