SASDE48: Toxin GraT (Putative killer protein, Toxin GraT., GraT)

Basic information

Entry

Database: SASBDB / ID: SASDE48

Sample

Toxin GraT

• Putative killer protein, Toxin GraT. (protein), GraT, Pseudomonas putida

• Function / homology: Toxin HigB-1 / RelE-like toxin of type II toxin-antitoxin system HigB / Toxin-antitoxin system, RelE/ParE toxin domain superfamily / molecular function activator activity / Putative Killer protein
• Biological species: Pseudomonas putida (bacteria)
• Citation: Journal: Nat Commun / Year: 2019; Title: A dual role in regulation and toxicity for the disordered N-terminus of the toxin GraT.; Authors: Ariel Talavera / Hedvig Tamman / Andres Ainelo / Albert Konijnenberg / San Hadži / Frank Sobott / Abel Garcia-Pino / Rita Hõrak / Remy Loris / ; Abstract: Bacterial toxin-antitoxin (TA) modules are tightly regulated to maintain growth in favorable conditions or growth arrest during stress. A typical regulatory strategy involves the antitoxin binding and repressing its own promoter while the toxin often acts as a co-repressor. Here we show that Pseudomonas putida graTA-encoded antitoxin GraA and toxin GraT differ from other TA proteins in the sense that not the antitoxin but the toxin possesses a flexible region. GraA auto-represses the graTA promoter: two GraA dimers bind cooperatively at opposite sides of the operator sequence. Contrary to other TA modules, GraT is a de-repressor of the graTA promoter as its N-terminal disordered segment prevents the binding of the GraTA complex to the operator. Removal of this region restores operator binding and abrogates Gr aT toxicity. GraTA represents a TA module where a flexible region in the toxin rather than in the antitoxin controls operon expression and toxin activity.

Contact author

• Ariel Talavera (VUB, Vrije Universiteit Brussel, Pleinlaan 2 1050 Brussel)

Models

Sample

• Sample: Name: Toxin GraT / Specimen concentration: 12.5 mg/ml
• Buffer: Name: 50 mM Tris 250 mM NaCl 2 mM TCEP / pH: 8

Entity #1379

Name: GraT / Type: protein / Description: Putative killer protein, Toxin GraT. / Formula weight: 11.104 / Num. of mol.: 1 / Source: Pseudomonas putida / References: UniProt: Q88MI5
Sequence:

MIRSFSCADT EALFTTGKTR RGSDIKSVAE RKLAMLDAAT ELRDLRSPPG NRLESLSGNR ADQHSIRVND QWRLCFTWTE HGPVNVEIVD YENLYFQ

Experimental information

• Beam: Instrument name: SOLEIL SWING / City: Saint-Aubin / 国: France / Type of source: X-ray synchrotronSynchrotron / Wavelength: 0.103317 Å / Dist. spec. to detc.: 1.499 mm
• Detector: Name: Eiger 4M / Pixsize x: 75 mm

Scan

Title: Toxin GraT / Measurement date: Sep 21, 2018 / Storage temperature: 24.8 °C / Cell temperature: 25 °C / Exposure time: 0.99 sec. / Number of frames: 40 / Unit: 1/A /

	Min	Max
Q	0.0146	0.6642

Distance distribution function P(R)

Sofotware P(R): GNOM 5.0 / Number of points: 825 /

	Min	Max
Q	0.0273667	0.526995
P(R) point	1	825
R	0	54.66

Result

Type of curve: sec /

	Experimental	Porod
MW	11.1 kDa	-
Volume	-	18 nm3

	P(R)	P(R) error	Guinier	Guinier error
Forward scattering, I0	0.0143	2.0E-5	0.01421	1.5E-5
Radius of gyration, Rg	1.55 nm	0.3	1.509 nm	0.03

	Min	Max
D	-	5.47
Guinier point	38	122