SASDEM4: HrpG/HrpV/HrpJ gatekeeper complex from Erwinia amylovora

Basic information

• Entry: Database: SASBDB / ID: SASDEM4

Sample

HrpG/HrpV/HrpJ gatekeeper complex from Erwinia amylovora

• Type III secretion protein HrpGType three secretion system (protein), Erwinia amylovora (strain CFBP1430)
• Type III secretion protein HrpVType three secretion system (protein), Erwinia amylovora (strain CFBP1430)
• Hypersensitivity response secretion protein HrpJ (protein), Erwinia amylovora (strain CFBP1430)

Function / homology

Function:

protein secretion by the type III secretion system / outer membrane / : / negative regulation of protein secretion / cell surface

Domain/homology:

TyeA superfamily / Type III secretion system effector delivery regulator TyeA-related / Type III secretion regulator, YopN/LcrE/InvE/MxiC / Hypersensitivity response secretion-like, HrpJ / HrpJ-like domain

Component:

Hypersensitivity response secretion protein HrpJ / Type III secretion protein HrpG / Type III secretion protein HrpV

• Biological species: Erwinia amylovora (strain CFBP1430) (bacteria)
• Citation: Journal: mBio / Year: 2018; Title: Migration of Type III Secretion System Transcriptional Regulators Links Gene Expression to Secretion.; Authors: Spyridoula N Charova / Anastasia D Gazi / Efstratios Mylonas / Charalambos Pozidis / Blanca Sabarit / Dimitrios Anagnostou / Konstantina Psatha / Michalis Aivaliotis / Carmen R Beuzon / Nickolas J Panopoulos / Michael Kokkinidis / ; Abstract: Many plant-pathogenic bacteria of considerable economic importance rely on type III secretion systems (T3SSs) of the Hrc-Hrp 1 family to subvert their plant hosts. T3SS gene expression is regulated through the HrpG and HrpV proteins, while secretion is controlled by the gatekeeper HrpJ. A link between the two mechanisms was so far unknown. Here, we show that a mechanistic coupling exists between the expression and secretion cascades through the direct binding of the HrpG/HrpV heterodimer, acting as a T3SS chaperone, to HrpJ. The ternary complex is docked to the cytoplasmic side of the inner bacterial membrane and orchestrates intermediate substrate secretion, without affecting early substrate secretion. The anchoring of the ternary complex to the membranes potentially keeps HrpG/HrpV away from DNA. In their multiple roles as transcriptional regulators and gatekeeper chaperones, HrpV/HrpG provide along with HrpJ potentially attractive targets for antibacterial strategies. On the basis of scientific/economic importance, and are considered among the top 10 plant-pathogenic bacteria in molecular plant pathology. Both employ type III secretion systems (T3SSs) of the Hrc-Hrp 1 family to subvert their plant hosts. For Hrc-Hrp 1, no functional link was known between the key processes of T3SS gene expression and secretion. Here, we show that a mechanistic coupling exists between expression and secretion cascades, through formation of a ternary complex involving the T3SS proteins HrpG, HrpV, and HrpJ. Our results highlight the functional and structural properties of a hitherto-unknown complex which orchestrates intermediate T3SS substrate secretion and may lead to better pathogen control through novel targets for antibacterial strategies.

Contact author

• Efstratios Mylonas (Institute of Molecular Biology and Biotechnology, FORTH)

Models

• Model #2378: ; Type: mix / Radius of dummy atoms: 1.90 A / Comment: CORAL model with reconstructed polypetide chain. / Chi-square value: 6.615184; Search similar-shape structures of this assembly by Omokage search (details)

Sample

• Sample: Name: HrpG/HrpV/HrpJ gatekeeper complex from Erwinia amylovora; Specimen concentration: 1.5 mg/ml / Entity id: 1296 / 1297 / 1298
• Buffer: Name: 20mM Tris-Cl, pH 8.0, 100mM NaCl, 2mM DTT, 0.5mM EDTA / pH: 8

Entity #1296

Type: protein
Description: Type III secretion protein HrpGType three secretion system
Formula weight: 19.166 / Source: Erwinia amylovora (strain CFBP1430) / References: UniProt: D4HVP1
Sequence:

MSYYHHHHHH DYDIPTTENL YFQGAMRSTE LQQWAQRWLD DVGTDQQLTV DDGVVWLQRR ADRFFALAEL TINASLDDEL LGRALQLSAP TLRYFGRDAA ALAQQGNSLI LALAIRQLEV NAVCQQLESL LNQRDVWQTM LQQPRRKAAT HGAVPLHSLA FLPRGNHG

Entity #1297

Type: protein
Description: Type III secretion protein HrpVType three secretion system
Formula weight: 13.164 / Source: Erwinia amylovora (strain CFBP1430) / References: UniProt: D4HVP4
Sequence:

MNNPCPQVTS LSALLSLLQL RQSCRWPVQQ GVELLALCGE RGRELMLNLQ PAFQPSGLYQ RLLSRRAQQL EIYDGCYLCL NHDRVLSCWR QLPESAMSDK QHIAQLFSLA GIHLAD

Entity #1298

Type: protein
Description: Hypersensitivity response secretion protein HrpJ
Formula weight: 42.4 / Source: Erwinia amylovora (strain CFBP1430) / References: UniProt: D4HVM8
Sequence:

MKIAPVLPGI QPPHPVAINA ADEAPAPTSV APSAAASASA RMPVSPLEDS MEEVSMTFAE HVERKSKALN QRRVVAQPGG RAQQHIERIE KLDELLKLLE HPQHARLETQ NQKMQTLLQH SASPGMDELL HAAGGDPARC DVLLRHTLLQ AQRTQNSELA TKAAQSLELL HKEKGAEVSA GLNTASAIAS FSTDPTQKQV MRQLYYQTIV HLQSGNAMLD ALLNSFGSAH FNQGLRTLQR ALADDIASRS SSIPRRALQK ILSSLQDASN ISQTLTASKT LLTRLAGKLP TINFTSLELT RRLLNLSGNG AYLRDLQNLS RDVAGQHPHH QVLFLSGLVP LVYHLPLGLW RDDKSKNRQM ALNLLRKVIS DYAGQEPRAS LTQPRTAL

Experimental information

• Beam: Instrument name: SOLEIL SWING / City: Saint-Aubin / 国: France / Type of source: X-ray synchrotronSynchrotron / Wavelength: 0.1033 Å / Dist. spec. to detc.: 3.1 mm
• Detector: Name: AVIEX PCCD170170 / Type: CCD

Scan

Title: HrpG/HrpV/HrpV gatekeeper complex from Erwinia amylovora
Measurement date: Apr 3, 2016 / Storage temperature: 4 °C / Cell temperature: 15 °C / Exposure time: 0.5 sec. / Number of frames: 30 / Unit: 1/A /

	Min	Max
Q	0.0216	0.4918

Distance distribution function P(R)

Sofotware P(R): GNOM 4.6 / Number of points: 456 /

	Min	Max
Q	0.02158	0.2798
P(R) point	1	456
R	0	160

Result

D max: 16 / Type of curve: single_conc /

	Experimental	Standard	Porod
MW	74.7 kDa	75 kDa	78 kDa

	P(R)	Guinier	Guinier error
Forward scattering, I0	0.2855	0.272	-
Radius of gyration, Rg	4.36 nm	4 nm	0.2