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Wolf-Watz, Hans
Alternative names
Publications (10 of 73) Show all publications
Ho, O., Rogne, P., Edgren, T., Wolf-Watz, H., Login, F. & Wolf-Watz, M. (2017). Characterization of the Ruler Protein Interaction Interface on the Substrate Specificity SwitchProtein in the Yersinia Type III Secretion System. Journal of Biological Chemistry, 292(8), 3299-3311
Open this publication in new window or tab >>Characterization of the Ruler Protein Interaction Interface on the Substrate Specificity SwitchProtein in the Yersinia Type III Secretion System
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2017 (English)In: Journal of Biological Chemistry, ISSN 0021-9258, E-ISSN 1083-351X, Vol. 292, no 8, p. 3299-3311Article, review/survey (Refereed) Published
Abstract [en]

Many pathogenic Gram-negative bacteria use the type III secretion system (T3SS) to deliver effector proteins into eukaryotic host cells. In Yersinia the switch to secretion of effector proteins is induced first after that intimate contact between the bacterium and its eukaryotic targetcell has been established and the T3SS proteins YscP and YscU are playing a central role in thisprocess. Here we identify the molecular details of the YscP binding site on YscU by means o fnuclear magnetic resonance (NMR) spectroscopy. The binding interface is centeredon the C-terminal domain of YscU. Disruptingthe YscU/YscP interaction by introducing point mutations at the interaction interface significantly reduced the secretion of effector proteins and HeLa cell cytotoxicity. Interestingly, the bindingof YscP to the slowly self-cleaving YscU variantP264A conferred significant protection againstauto-proteolysis. The YscP mediated inhibition of YscU auto-proteolysis suggest that the cleavage event may act as a timing switch in the regulationof early vs. late T3SS substrates. We also show that YscUC binds to the inner-rod protein YscI with a Kd of 3.8 μM and with one-to-one stoichiometry. The significant similarity between different members of the YscU, YscP, YscI families suggests that the protein-protein interactions discussed in this study are alsorelevant for other T3SS-containing Gram-negative bacteria.

National Category
Biochemistry and Molecular Biology
Research subject
Biochemistry
Identifiers
urn:nbn:se:umu:diva-130048 (URN)10.1074/jbc.M116.770255 (DOI)000395538800021 ()
Available from: 2017-01-11 Created: 2017-01-11 Last updated: 2018-06-09Bibliographically approved
Gupta, A., Reinartz, I., Spilotros, A., Jonna, V. R., Hofer, A., Svergun, D. I., . . . Wolf-Watz, M. (2017). Global Disordering in Stereo-Specific Protein Association. Paper presented at 61st Annual Meeting of the Biophysical-Society, FEB 11-15, 2017, New Orleans, LA. Biophysical Journal, 112(3), 33A-33A
Open this publication in new window or tab >>Global Disordering in Stereo-Specific Protein Association
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2017 (English)In: Biophysical Journal, ISSN 0006-3495, E-ISSN 1542-0086, Vol. 112, no 3, p. 33A-33AArticle in journal, Meeting abstract (Refereed) Published
Place, publisher, year, edition, pages
CELL PRESS, 2017
National Category
Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)
Identifiers
urn:nbn:se:umu:diva-137015 (URN)000402328000166 ()
Conference
61st Annual Meeting of the Biophysical-Society, FEB 11-15, 2017, New Orleans, LA
Available from: 2017-06-29 Created: 2017-06-29 Last updated: 2018-06-09Bibliographically approved
Wang, H., Avican, K., Fahlgren, A., Erttmann, S. F., Nuss, A. M., Dersch, P., . . . Wolf-Watz, H. (2016). Increased plasmid copy number is essential for Yersinia T3SS function and virulence. Science, 353(6298), 492-495
Open this publication in new window or tab >>Increased plasmid copy number is essential for Yersinia T3SS function and virulence
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2016 (English)In: Science, ISSN 0036-8075, E-ISSN 1095-9203, Vol. 353, no 6298, p. 492-495Article in journal (Refereed) Published
Abstract [en]

Pathogenic bacteria have evolved numerous virulence mechanisms that are essential for establishing infections. The enterobacterium Yersinia uses a type III secretion system (T3SS) encoded by a 70-kilobase, low-copy, IncFII-class virulence plasmid. We report a novel virulence strategy in Y. pseudotuberculosis in which this pathogen up-regulates the plasmid copy number during infection. We found that an increased dose of plasmid-encoded genes is indispensable for virulence and substantially elevates the expression and function of the T3SS. Remarkably, we observed direct, tight coupling between plasmid replication and T3SS function. This regulatory pathway provides a framework for further exploration of the environmental sensing mechanisms of pathogenic bacteria.

National Category
Microbiology in the medical area Infectious Medicine
Identifiers
urn:nbn:se:umu:diva-125586 (URN)10.1126/science.aaf7501 (DOI)000380583600042 ()27365311 (PubMedID)
Available from: 2016-09-19 Created: 2016-09-13 Last updated: 2018-06-07Bibliographically approved
Engström, P., Krishnan, K. S., Ngyuen, B. D., Chorell, E., Normark, J., Silver, J., . . . Bergström, S. (2015). A 2-Pyridone-Amide Inhibitor Targets the Glucose Metabolism Pathway of Chlamydia trachomatis. mBio, 6(1), Article ID e02304-14.
Open this publication in new window or tab >>A 2-Pyridone-Amide Inhibitor Targets the Glucose Metabolism Pathway of Chlamydia trachomatis
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2015 (English)In: mBio, ISSN 2161-2129, E-ISSN 2150-7511, Vol. 6, no 1, article id e02304-14Article in journal (Refereed) Published
Abstract [en]

In a screen for compounds that inhibit infectivity of the obligate intracellular pathogen Chlamydia trachomatis, we identified the 2-pyridone amide KSK120. A fluorescent KSK120 analogue was synthesized and observed to be associated with the C. trachomatis surface, suggesting that its target is bacterial. We isolated KSK120-resistant strains and determined that several resistance mutations are in genes that affect the uptake and use of glucose-6-phosphate (G-6P). Consistent with an effect on G-6P metabolism, treatment with KSK120 blocked glycogen accumulation. Interestingly, KSK120 did not affect Escherichia coli or the host cell. Thus, 2-pyridone amides may represent a class of drugs that can specifically inhibit C. trachomatis infection. IMPORTANCE Chlamydia trachomatis is a bacterial pathogen of humans that causes a common sexually transmitted disease as well as eye infections. It grows only inside cells of its host organism, within a parasitophorous vacuole termed the inclusion. Little is known, however, about what bacterial components and processes are important for C. trachomatis cellular infectivity. Here, by using a visual screen for compounds that affect bacterial distribution within the chlamydial inclusion, we identified the inhibitor KSK120. As hypothesized, the altered bacterial distribution induced by KSK120 correlated with a block in C. trachomatis infectivity. Our data suggest that the compound targets the glucose-6-phosphate (G-6P) metabolism pathway of C. trachomatis, supporting previous indications that G-6P metabolism is critical for C. trachomatis infectivity. Thus, KSK120 may be a useful tool to study chlamydial glucose metabolism and has the potential to be used in the treatment of C. trachomatis infections.

National Category
Microbiology in the medical area
Identifiers
urn:nbn:se:umu:diva-102254 (URN)10.1128/mBio.02304-14 (DOI)000350631900021 ()25550323 (PubMedID)
Available from: 2015-04-22 Created: 2015-04-22 Last updated: 2018-06-07Bibliographically approved
Login, F. H. & Wolf-Watz, H. (2015). YscU/FlhB of Yersinia pseudotuberculosis Harbors a C-terminal Type III Secretion Signal. Journal of Biological Chemistry, 290(43), 26282-26291
Open this publication in new window or tab >>YscU/FlhB of Yersinia pseudotuberculosis Harbors a C-terminal Type III Secretion Signal
2015 (English)In: Journal of Biological Chemistry, ISSN 0021-9258, E-ISSN 1083-351X, Vol. 290, no 43, p. 26282-26291Article in journal (Refereed) Published
Abstract [en]

All type III secretion systems (T3SS) harbor a member of the YscU/FlhB family of proteins that is characterized by an auto-proteolytic process that occurs at a conserved cytoplasmic NPTH motif. We have previously demonstrated that YscU(CC), the C-terminal peptide generated by auto-proteolysis of Yersinia pseudotuberculosis YscU, is secreted by the T3SS when bacteria are grown in Ca2+ -depleted medium at 37 degrees C. Here, we investigated the secretion of this early T3S-substrate and showed that YscU(CC) encompasses a specific C-terminal T3S signal within the 15 last residues (U-15). U-15 promoted C-terminal secretion of reporter proteins like GST and YopE lacking its native secretion signal. Similar to the "classical" N-terminal secretion signal, U-15 interacted with the ATPase YscN. Although U-15 is critical for YscU(CC) secretion, deletion of the C-terminal secretion signal of YscU(CC) did neither affect Yop secretion nor Yop translocation. However, these deletions resulted in increased secretion of YscF, the needle subunit. Thus, these results suggest that YscU via its C-terminal secretion signal is involved in regulation of the YscF secretion.

Place, publisher, year, edition, pages
Umea Univ, Dept Mol Biol, Umea Ctr Microbial Res, SE-90187 Umea, Sweden. Umea Univ, Lab Mol Infect Med Sweden MIMS, Umea Ctr Microbial Res, SE-90187 Umea, Sweden.: , 2015
National Category
Microbiology in the medical area
Identifiers
urn:nbn:se:umu:diva-111762 (URN)10.1074/jbc.M114.633677 (DOI)000363527300044 ()26338709 (PubMedID)
Available from: 2015-11-25 Created: 2015-11-23 Last updated: 2018-06-07Bibliographically approved
Weise, C., Login, F. H., Ho, O., Gröbner, G., Wolf-Watz, H. & Wolf-Watz, M. (2014). Negatively charged lipid membranes promote a disorder-order transition in the Yersinia YscU protein. Biophysical Journal, 107(8), 1950-1961
Open this publication in new window or tab >>Negatively charged lipid membranes promote a disorder-order transition in the Yersinia YscU protein
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2014 (English)In: Biophysical Journal, ISSN 0006-3495, E-ISSN 1542-0086, Vol. 107, no 8, p. 1950-1961Article in journal (Refereed) Published
Abstract [en]

The inner membrane of Gram-negative bacteria is negatively charged, rendering positively charged cytoplasmic proteins in close proximity likely candidates for protein-membrane interactions. YscU is a Yersinia pseudotuberculosis type III secretion system protein crucial for bacterial pathogenesis. The protein contains a highly conserved positively charged linker sequence that separates membrane-spanning and cytoplasmic (YscUC) domains. Although disordered in solution, inspection of the primary sequence of the linker reveals that positively charged residues are separated with a typical helical periodicity. Here, we demonstrate that the linker sequence of YscU undergoes a largely electrostatically driven coil-to-helix transition upon binding to negatively charged membrane interfaces. Using membrane-mimicking sodium dodecyl sulfate micelles, an NMR derived structural model reveals the induction of three helical segments in the linker. The overall linker placement in sodium dodecyl sulfate micelles was identified by NMR experiments including paramagnetic relaxation enhancements. Partitioning of individual residues agrees with their hydrophobicity and supports an interfacial positioning of the helices. Replacement of positively charged linker residues with alanine resulted in YscUC variants displaying attenuated membrane-binding affinities, suggesting that the membrane interaction depends on positive charges within the linker. In vivo experiments with bacteria expressing these YscU replacements resulted in phenotypes displaying significantly reduced effector protein secretion levels. Taken together, our data identify a previously unknown membrane-interacting surface of YscUC that, when perturbed by mutations, disrupts the function of the pathogenic machinery in Yersinia.

Place, publisher, year, edition, pages
Cell Press, 2014
National Category
Biophysics
Identifiers
urn:nbn:se:umu:diva-95192 (URN)10.1016/j.bpj.2014.09.005 (DOI)000343682700021 ()25418176 (PubMedID)
Available from: 2014-10-23 Created: 2014-10-23 Last updated: 2018-06-07Bibliographically approved
Antti, H., Fahlgren, A., Näsström, E., Kouremenos, K., Sundén-Cullberg, J., Guo, Y., . . . Fällman, M. (2013). Metabolic profiling for detection of staphylococcus aureus infection and antibiotic resistance. PLoS ONE, 8(2), Article ID e56971.
Open this publication in new window or tab >>Metabolic profiling for detection of staphylococcus aureus infection and antibiotic resistance
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2013 (English)In: PLoS ONE, ISSN 1932-6203, E-ISSN 1932-6203, Vol. 8, no 2, article id e56971Article in journal (Refereed) Published
Abstract [en]

Due to slow diagnostics, physicians must optimize antibiotic therapies based on clinical evaluation of patients without specific information on causative bacteria. We have investigated metabolomic analysis of blood for the detection of acute bacterial infection and early differentiation between ineffective and effective antibiotic treatment. A vital and timely therapeutic difficulty was thereby addressed: the ability to rapidly detect treatment failures because of antibiotic-resistant bacteria. Methicillin-resistant (MRSA) and methicillin-sensitive (MSSA) were used and for infecting mice, while natural MSSA infection was studied in humans. Samples of bacterial growth media, the blood of infected mice and of humans were analyzed with combined Gas Chromatography/Mass Spectrometry. Multivariate data analysis was used to reveal the metabolic profiles of infection and the responses to different antibiotic treatments. experiments resulted in the detection of 256 putative metabolites and mice infection experiments resulted in the detection of 474 putative metabolites. Importantly, ineffective and effective antibiotic treatments were differentiated already two hours after treatment start in both experimental systems. That is, the ineffective treatment of MRSA using cloxacillin and untreated controls produced one metabolic profile while all effective treatment combinations using cloxacillin or vancomycin for MSSA or MRSA produced another profile. For further evaluation of the concept, blood samples of humans admitted to intensive care with severe sepsis were analyzed. One hundred thirty-three putative metabolites differentiated severe MSSA sepsis (n = 6) from severe sepsis (n = 10) and identified treatment responses over time. Combined analysis of human, , and mice samples identified 25 metabolites indicative of effective treatment of sepsis. Taken together, this study provides a proof of concept of the utility of analyzing metabolite patterns in blood for early differentiation between ineffective and effective antibiotic treatment in acute infections.

National Category
Microbiology in the medical area Infectious Medicine
Identifiers
urn:nbn:se:umu:diva-66816 (URN)10.1371/journal.pone.0056971 (DOI)000316849500048 ()23451124 (PubMedID)
Available from: 2013-03-05 Created: 2013-03-05 Last updated: 2018-06-08Bibliographically approved
Engström, P., Nguyen, B. D., Normark, J., Nilsson, I., Bastidas, R. J., Gylfe, Å., . . . Bergström, S. (2013). Mutations in hemG Mediate Resistance to Salicylidene Acylhydrazides, Demonstrating a Novel Link between Protoporphyrinogen Oxidase (HemG) and Chlamydia trachomatis Infectivity. Journal of Bacteriology, 195(18), 4221-4230
Open this publication in new window or tab >>Mutations in hemG Mediate Resistance to Salicylidene Acylhydrazides, Demonstrating a Novel Link between Protoporphyrinogen Oxidase (HemG) and Chlamydia trachomatis Infectivity
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2013 (English)In: Journal of Bacteriology, ISSN 0021-9193, E-ISSN 1098-5530, Vol. 195, no 18, p. 4221-4230Article in journal (Refereed) Published
Abstract [en]

Salicylidene acylhydrazides (SAHs) inhibit the type III secretion system (T3S) of Yersinia and other Gram-negative bacteria. In addition, SAHs restrict the growth and development of Chlamydia species. However, since the inhibition of Chlamydia growth by SAH is suppressed by the addition of excess iron and since SAHs have an iron-chelating capacity, their role as specific T3S inhibitors is unclear. We investigated here whether SAHs exhibit a function on C. trachomatis that goes beyond iron chelation. We found that the iron-saturated SAH INP0341 (IS-INP0341) specifically affects C. trachomatis infectivity with reduced generation of infectious elementary body (EB) progeny. Selection and isolation of spontaneous SAH-resistant mutant strains revealed that mutations in hemG suppressed the reduced infectivity caused by IS-INP0341 treatment. Structural modeling of C. trachomatis HemG predicts that the acquired mutations are located in the active site of the enzyme, suggesting that IS-INP0341 inhibits this domain of HemG and that protoporphyrinogen oxidase (HemG) and heme metabolism are important for C. trachomatis infectivity.

Place, publisher, year, edition, pages
Washington DC, USA: American Society for Microbiology, 2013
National Category
Biochemistry and Molecular Biology
Identifiers
urn:nbn:se:umu:diva-80746 (URN)10.1128/JB.00506-13 (DOI)000323649300023 ()
Available from: 2013-10-07 Created: 2013-09-25 Last updated: 2018-06-08Bibliographically approved
Costa, T., Amer, A., Farag, S., Wolf-Watz, H., Fällman, M., Fahlgren, A., . . . Francis, M. (2013). Type III secretion translocon assemblies that attenuate Yersinia virulence. Cellular Microbiology, 15(7), 1088-1110
Open this publication in new window or tab >>Type III secretion translocon assemblies that attenuate Yersinia virulence
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2013 (English)In: Cellular Microbiology, ISSN 1462-5814, E-ISSN 1462-5822, Vol. 15, no 7, p. 1088-1110Article in journal (Refereed) Published
Abstract [en]

Type III secretion enables bacteria to intoxicate eukaryotic cells with anti-host effectors. A class of secreted cargo are the two hydrophobic translocators that form a translocon pore in the host cell plasma membrane through which the translocated effectors may gain cellular entry. In pathogenic Yersinia, YopB and YopD shape this translocon pore. Here, four in cis yopD mutations were constructed to disrupt a predicted α-helix motif at the C-terminus. Mutants YopD(I262P) and YopD(K267P) poorly localized Yop effectors into target eukaryotic cells and failed to resist uptake and killing by immune cells. These defects were due to deficiencies in host-membrane insertion of the YopD-YopB translocon. Mutants YopD(A263P) and YopD(A270P) had no measurable in vitro translocation defect, even though they formed smaller translocon pores in erythrocyte membranes. Despite this, all four mutants were attenuated in a mouse infection model. Hence, YopD variants have been generated that can spawn translocons capable of targeting effectors in vitro, yet were bereft of any lethal effect in vivo. Therefore, Yop translocators may possess other in vivo functions that extend beyond being a portal for effector delivery into host cells.

Place, publisher, year, edition, pages
Wiley-Blackwell, 2013
Keywords
virulence, coiled-coil, effector delivery, pore formation, regulation, YopD
National Category
Microbiology Biochemistry and Molecular Biology Microbiology in the medical area
Research subject
Infectious Diseases; Microbiology; Molecular Biology
Identifiers
urn:nbn:se:umu:diva-67117 (URN)10.1111/cmi.12100 (DOI)000320394800004 ()
Funder
Swedish Research Council
Note

This work, performed within the framework of the Umea Centre for Microbial Research-Linnaeus Program, was supported by grants from the Carl Tryggers Foundation for Scientific Research (M. S. F.), Swedish Research Council (H.W.-W., M. F., T. E., M. S. F.), Foundation for Medical Research at Umea University (M. S. F.) and J C Kempe Memorial Fund (T. R. C., A. A. A.).

Available from: 2013-03-13 Created: 2013-03-13 Last updated: 2018-06-08Bibliographically approved
Frost, S., Ho, O., Login, F. H., Weise, C. F., Wolf-Watz, H. & Wolf-Watz, M. (2012). Autoproteolysis and Intramolecular Dissociation of Yersinia YscU Precedes Secretion of Its C-Terminal Polypeptide YscU CC. PLoS ONE, 7(11), Article ID e49349.
Open this publication in new window or tab >>Autoproteolysis and Intramolecular Dissociation of Yersinia YscU Precedes Secretion of Its C-Terminal Polypeptide YscU CC
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2012 (English)In: PLoS ONE, ISSN 1932-6203, E-ISSN 1932-6203, Vol. 7, no 11, article id e49349Article in journal (Refereed) Published
Abstract [en]

Type III secretion system mediated secretion and translocation of Yop-effector proteins across the eukaryotic target cell membrane by pathogenic Yersinia is highly organized and is dependent on a switching event from secretion of early structural substrates to late effector substrates (Yops). Substrate switching can be mimicked in vitro by modulating the calcium levels in the growth medium. YscU that is essential for regulation of this switch undergoes autoproteolysis at a conserved N↑PTH motif, resulting in a 10 kDa C-terminal polypeptide fragment denoted YscUCC. Here we show that depletion of calcium induces intramolecular dissociation of YscUCC from YscU followed by secretion of the YscUCC polypeptide. Thus, YscUCC behaved in vivo as a Yop protein with respect to secretion properties. Further, destabilized yscU mutants displayed increased rates of dissociation of YscUCC in vitro resulting in enhanced Yop secretion in vivo at 30°C relative to the wild-type strain.These findings provide strong support to the relevance of YscUCC dissociation for Yop secretion. We propose that YscUCC orchestrates a block in the secretion channel that is eliminated by calcium depletion. Further, the striking homology between different members of the YscU/FlhB family suggests that this protein family possess regulatory functions also in other bacteria using comparable mechanisms.

National Category
Chemical Sciences
Identifiers
urn:nbn:se:umu:diva-61703 (URN)10.1371/journal.pone.0049349 (DOI)000311821000040 ()23185318 (PubMedID)
Available from: 2012-11-23 Created: 2012-11-23 Last updated: 2018-06-08Bibliographically approved
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