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Lavander, Moa
Publications (10 of 14) Show all publications
Avican, U., Beckstette, M., Heroven, A. K., Lavander, M., Dersch, P. & Forsberg, Å. (2016). Transcriptomic and phenotypic analysis reveals new functions for the Tat pathway in Yersinia pseudotuberculosis. Journal of Bacteriology, 198(20), 2876-2886
Open this publication in new window or tab >>Transcriptomic and phenotypic analysis reveals new functions for the Tat pathway in Yersinia pseudotuberculosis
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2016 (English)In: Journal of Bacteriology, ISSN 0021-9193, E-ISSN 1098-5530, Vol. 198, no 20, p. 2876-2886Article in journal (Refereed) Published
Abstract [en]

The Twin-arginine translocation (Tat) system mediates secretion of folded proteins that in bacteria, plants and archaea are identified via an N-terminal signal peptide. Tat systems are associated with virulence in many bacterial pathogens and our previous studies revealed that Tat deficient Yersinia pseudotuberculosis was severely attenuated for virulence. Aiming to identify Tat-dependent pathways and phenotypes of relevance for in vivo infection, we analysed the global transcriptome of parental and ∆tatC mutant strains of Y. pseudotuberculosis during exponential and stationary growth at 26oC and 37oC. The most significant changes in the transcriptome of the ∆tatC mutant were seen at 26oC during stationary phase growth and these included the altered expression of genes related to virulence, stress responses and metabolism. Subsequent phenotypic analysis based on these transcriptome changes revealed several novel Tat-dependent phenotypes including decreased YadA expression, impaired growth under iron-limiting and high copper conditions as well as acidic pH and SDS. Several functionally related Tat substrates were also verified to contribute to these phenotypes. Interestingly, the phenotypic defects observed in the Tat-deficient strain were generally more pronounced than in mutants lacking the Tat substrate predicted to contribute to that specific function. Altogether, this provides new insight into the impact of Tat deficiency on in vivo fitness and survival/replication of Y. pseudotuberculosis during infection.

Place, publisher, year, edition, pages
Washington: American Society for Microbiology, 2016
Keywords
Yersinia pseudotuberculosis, Tat pathway, virulence, stress response, transcriptome analysis
National Category
Microbiology Bioinformatics and Systems Biology Biochemistry and Molecular Biology
Research subject
Infectious Diseases; Microbiology; Molecular Biology
Identifiers
urn:nbn:se:umu:diva-128029 (URN)10.1128/JB.00352-16 (DOI)000384347500014 ()2-s2.0-84991200249 (Scopus ID)
Available from: 2016-11-22 Created: 2016-11-22 Last updated: 2023-03-23Bibliographically approved
Sun, K., Bröms, J., Lavander, M., Gurram, B. K., Enquist, P.-A., Andersson, C. D., . . . Sjöstedt, A. (2014). Screening for inhibition of Vibrio cholerae VipA-VipB interaction identifies small-molecule compounds active against type VI secretion. Antimicrobial Agents and Chemotherapy, 58(7), 4123-4130
Open this publication in new window or tab >>Screening for inhibition of Vibrio cholerae VipA-VipB interaction identifies small-molecule compounds active against type VI secretion
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2014 (English)In: Antimicrobial Agents and Chemotherapy, ISSN 0066-4804, E-ISSN 1098-6596, Vol. 58, no 7, p. 4123-4130Article in journal (Refereed) Published
Abstract [en]

The type VI secretion system (T6SS) is the most prevalent bacterial secretion system and an important virulence mechanism utilized by Gram-negative bacteria, either to target eukaryotic cells or to combat other microbes. The components show much variability, but some appear essential for the function, and two homologues, denoted VipA and VipB in Vibrio cholerae, have been identified in all T6SSs described so far. Secretion is dependent on binding of an alpha-helical region of VipA to VipB, and in the absence of this binding, both components are degraded within minutes and secretion is ceased. The aim of the study was to investigate if this interaction could be blocked, and we hypothesized that such inhibition would lead to abrogation of T6S. A library of 9,600 small-molecule compounds was screened for their ability to block the binding of VipA-VipB in a bacterial two-hybrid system (B2H). After excluding compounds that showed cytotoxicity toward eukaryotic cells, that inhibited growth of Vibrio, or that inhibited an unrelated B2H interaction, 34 compounds were further investigated for effects on the T6SS-dependent secretion of hemolysin-coregulated protein (Hcp) or of phospholipase A(1) activity. Two compounds, KS100 and KS200, showed intermediate or strong effects in both assays. Analogues were obtained, and compounds with potent inhibitory effects in the assays and desirable physicochemical properties as predicted by in silico analysis were identified. Since the compounds specifically target a virulence mechanism without affecting bacterial replication, they have the potential to mitigate the virulence with minimal risk for development of resistance.

National Category
Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy) Pharmacology and Toxicology
Identifiers
urn:nbn:se:umu:diva-91844 (URN)10.1128/AAC.02819-13 (DOI)000338846500069 ()24798289 (PubMedID)2-s2.0-84903202590 (Scopus ID)
Available from: 2014-08-27 Created: 2014-08-18 Last updated: 2023-03-24Bibliographically approved
Bröms, J. E., Meyer, L., Lavander, M., Larsson, P. & Sjöstedt, A. (2012). DotU and VgrG, core components of type VI secretion systems, are essential for Francisella LVS pathogenicity. PLOS ONE, 7(4), Article ID e34639.
Open this publication in new window or tab >>DotU and VgrG, core components of type VI secretion systems, are essential for Francisella LVS pathogenicity
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2012 (English)In: PLOS ONE, E-ISSN 1932-6203, Vol. 7, no 4, article id e34639Article in journal (Refereed) Published
Abstract [en]

The Gram-negative bacterium Francisella tularensis causes tularemia, a disease which requires bacterial escape from phagosomes of infected macrophages. Once in the cytosol, the bacterium rapidly multiplies, inhibits activation of the inflammasome and ultimately causes death of the host cell. Of importance for these processes is a 33-kb gene cluster, the Francisella pathogenicity island (FPI), which is believed to encode a type VI secretion system (T6SS). In this study, we analyzed the role of the FPI-encoded proteins VgrG and DotU, which are conserved components of type VI secretion (T6S) clusters. We demonstrate that in F. tularensis LVS, VgrG was shown to form multimers, consistent with its suggested role as a trimeric membrane puncturing device in T6SSs, while the inner membrane protein DotU was shown to stabilize PdpB/IcmF, another T6SS core component. Upon infection of J774 cells, both Delta vgrG and Delta dotU mutants did not escape from phagosomes, and subsequently, did not multiply or cause cytopathogenicity. They also showed impaired activation of the inflammasome and marked attenuation in the mouse model. Moreover, all of the DotU-dependent functions investigated here required the presence of three residues that are essentially conserved among all DotU homologues. Thus, in agreement with a core function in T6S clusters, VgrG and DotU play key roles for modulation of the intracellular host response as well as for the virulence of F. tularensis.

National Category
Microbiology in the medical area
Identifiers
urn:nbn:se:umu:diva-57392 (URN)10.1371/journal.pone.0034639 (DOI)000305341600057 ()2-s2.0-84859724534 (Scopus ID)
Available from: 2012-07-17 Created: 2012-07-16 Last updated: 2023-03-24Bibliographically approved
Bröms, J. E., Meyer, L., Sun, K., Lavander, M. & Sjöstedt, A. (2012). Unique substrates secreted by the type VI secretion system of Francisella tularensis during intramacrophage infection. PLOS ONE, 7(11), Article ID e50473.
Open this publication in new window or tab >>Unique substrates secreted by the type VI secretion system of Francisella tularensis during intramacrophage infection
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2012 (English)In: PLOS ONE, E-ISSN 1932-6203, Vol. 7, no 11, article id e50473Article in journal (Refereed) Published
Abstract [en]

Gram-negative bacteria have evolved sophisticated secretion machineries specialized for the secretion of macromolecules important for their life cycles. The Type VI secretion system (T6SS) is the most widely spread bacterial secretion machinery and is encoded by large, variable gene clusters, often found to be essential for virulence. The latter is true for the atypical T6SS encoded by the Francisella pathogenicity island (FPI) of the highly pathogenic, intracellular bacterium Francisella tularensis. We here undertook a comprehensive analysis of the intramacrophage secretion of the 17 FPI proteins of the live vaccine strain, LVS, of F. tularensis. All were expressed as fusions to the TEM beta-lactamase and cleavage of the fluorescent substrate CCF2-AM, a direct consequence of the delivery of the proteins into the macrophage cytosol, was followed over time. The FPI proteins IglE, IglC, VgrG, IglI, PdpE, PdpA, IglJ and IglF were all secreted, which was dependent on the core components DotU, VgrG, and IglC, as well as IglG. In contrast, the method was not directly applicable on F. novicida U112, since it showed very intense native beta-lactamase secretion due to FTN_1072. Its role was proven by ectopic expression in trans in LVS. We did not observe secretion of any of the LVS substrates VgrG, IglJ, IglF or IglI, when tested in a FTN_1072 deficient strain of F. novicida, whereas IglE, IglC, PdpA and even more so PdpE were all secreted. This suggests that there may be fundamental differences in the T6S mechanism among the Francisella subspecies. The findings further corroborate the unusual nature of the T6SS of F. tularensis since almost all of the identified substrates are unique to the species.

National Category
Microbiology in the medical area
Identifiers
urn:nbn:se:umu:diva-63027 (URN)10.1371/journal.pone.0050473 (DOI)000311535700086 ()23185631 (PubMedID)2-s2.0-84869842318 (Scopus ID)
Available from: 2012-12-28 Created: 2012-12-27 Last updated: 2023-03-23Bibliographically approved
Bröms, J. E., Lavander, M., Meyer, L. & Sjöstedt, A. (2011). IglG and IglI of the Francisella pathogenicity island are important virulence determinants of Francisella tularensis LVS. Infection and Immunity, 79(9), 3683-3696
Open this publication in new window or tab >>IglG and IglI of the Francisella pathogenicity island are important virulence determinants of Francisella tularensis LVS
2011 (English)In: Infection and Immunity, ISSN 0019-9567, E-ISSN 1098-5522, Vol. 79, no 9, p. 3683-3696Article in journal (Refereed) Published
Abstract [en]

The Gram-negative bacterium Francisella tularensis is the causative agent of tularemia, a disease intimately associated with the multiplication of the bacterium within host macrophages. This in turn requires the expression of Francisella pathogenicity island (FPI) genes, believed to encode a type VI secretion system. While the exact functions of many of the components have yet to be revealed, some have been found to contribute to the ability of Francisella to cause systemic infection in mice as well as to prevent phagolysosomal fusion and facilitate escape into the host cytosol. Upon reaching this compartment, the bacterium rapidly multiplies, inhibits activation of the inflammasome, and ultimately causes apoptosis of the host cell. In this study, we analyzed the contribution of the FPI-encoded proteins IglG, IglI, and PdpE to the aforementioned processes in F. tularensis LVS. The ΔpdpE mutant behaved similarly to the parental strain in all investigated assays. In contrast, ΔiglG and ΔiglI mutants, although they were efficiently replicating in J774A.1 cells, both exhibited delayed phagosomal escape, conferred a delayed activation of the inflammasome, and exhibited reduced cytopathogenicity as well as marked attenuation in the mouse model. Thus, IglG and IglI play key roles for modulation of the intracellular host response and also for the virulence of F. tularensis.

Place, publisher, year, edition, pages
American Society for Microbiology, 2011
National Category
Immunology in the medical area Infectious Medicine
Identifiers
urn:nbn:se:umu:diva-83830 (URN)10.1128/IAI.01344-10 (DOI)000293891000021 ()21690239 (PubMedID)2-s2.0-80052323146 (Scopus ID)
Funder
Swedish Research Council, 2006-3426, 2006-2877, 2009-5026
Available from: 2013-12-09 Created: 2013-12-09 Last updated: 2023-03-23Bibliographically approved
Bröms, J. E., Lavander, M. & Sjöstedt, A. (2010). Dissection of the Functions of the IglC Protein of Francisella tularensis. In: The challenge of highly pathogenic microorganisms: mechanisms of virulence and novel medical countermeasures. Paper presented at 46th Oholo Conference on the Challenge of Highly Pathogenic Microorganisms - Mechanisms of Virulence and Novel Medical Countermeasures, Eilat, Israel, October 25-29, 2009 (pp. 67-75). Springer
Open this publication in new window or tab >>Dissection of the Functions of the IglC Protein of Francisella tularensis
2010 (English)In: The challenge of highly pathogenic microorganisms: mechanisms of virulence and novel medical countermeasures, Springer, 2010, p. 67-75Conference paper, Published paper (Refereed)
Abstract [en]

Francisella tularensis harbors genes with similarity to genes encoding components of a type VI secretion system (T6SS). These include iglA and iglB, the homologues of which are conserved in T6SSs. They are part of the igl operon, also encompassing the iglC and iglD genes. We have used a yeast two-hybrid system to study the interaction of the Igl proteins of E tularensis LVS. Previously, we identified a region of IglA necessary for efficient binding to IglB as well as for IglAB protein stability and intra-macrophage growth with an essential role for a conserved alpha-helical region. Thus, IglA-IglB complex formation is clearly crucial for Francisella pathogenicity and the same interaction is conserved in other human pathogens. Herein, the interaction of IglC with other members of the operon was investigated. It showed no binding to the other members in the yeast two-hybrid assay and we found also that two cysteine residues, C191 and C192, predicted to be putative prenylation sites, played no role for the important contribution of IglC to the intracellular replication of E tularensis although C191 was important for the stability of the protein.

Place, publisher, year, edition, pages
Springer, 2010
Keywords
Francisella, IglC, Yeast two-hybrid system, Cysteine residues
National Category
Infectious Medicine Microbiology in the medical area
Identifiers
urn:nbn:se:umu:diva-109871 (URN)10.1007/978-90-481-9054-6_7 (DOI)000322847000007 ()978-90-481-9053-9 (ISBN)978-90-481-9054-6 (e-book) (ISBN)
Conference
46th Oholo Conference on the Challenge of Highly Pathogenic Microorganisms - Mechanisms of Virulence and Novel Medical Countermeasures, Eilat, Israel, October 25-29, 2009
Available from: 2015-10-07 Created: 2015-10-07 Last updated: 2018-06-07Bibliographically approved
Bröms, J. E., Lavander, M. & Sjöstedt, A. (2009). A conserved α-helix essential for a type VI secretion-like system of Francisella tularensis. Journal of Bacteriology, 191(8), 2431-2446
Open this publication in new window or tab >>A conserved α-helix essential for a type VI secretion-like system of Francisella tularensis
2009 (English)In: Journal of Bacteriology, ISSN 0021-9193, E-ISSN 1098-5530, Vol. 191, no 8, p. 2431-2446Article in journal (Refereed) Published
Abstract [en]

Francisella tularensis harbors genes with similarity to genes encoding components of a type VI secretion system (T6SS) recently identified in several gram-negative bacteria. These include iglA and iglB, the homologues of which are conserved in most T6SSs. We used a yeast two-hybrid system to study the interaction of the Igl proteins of F. tularensis LVS. We identified a region of IglA, encompassing residues 33-132, necessary for efficient binding to IglB as well as for IglAB protein stability and intra-macrophage growth. In particular, residues 103-122, overlapping with a highly conserved alpha-helix, played an absolutely essential role. Point mutations within this domain caused modest defects in IglA-IglB binding in yeast, but markedly impaired intra-macrophage replication and phagosomal escape, resulting in severe attenuation of LVS in mice. Thus, IglA-IglB complex formation is clearly crucial for Francisella pathogenicity. This interaction may be universal to T6S, since IglAB homologues of Yersinia pseudotuberculosis, Pseudomonas aeruginosa, Vibrio cholerae, Salmonella typhimurium and Escherichia coli were also shown to interact in yeast and the interaction was dependent on the preservation of the same alpha-helix. Heterologous interactions formed between non-native IglAB proteins further supported the notion of a conserved binding site. Thus, IglA-IglB complex formation is clearly crucial for Francisella pathogenicity and the same interaction is conserved in other human pathogens.

Keywords
pathogenicity island; protein secretion; escherichia-coli; yersinia-pestis; pseudomonas-aeruginosa; edwardsiella-tarda; virulence factors; vibrio-cholerae; live vaccine; escape
National Category
Microbiology
Identifiers
urn:nbn:se:umu:diva-19549 (URN)10.1128/JB.01759-08 (DOI)19201795 (PubMedID)2-s2.0-65249149518 (Scopus ID)
Available from: 2009-03-06 Created: 2009-03-06 Last updated: 2023-03-24Bibliographically approved
Björnfot, A.-C., Lavander, M., Forsberg, Å. & Wolf-Watz, H. (2009). Autoproteolysis of YscU of Yersinia pseudotuberculosis is important for regulation of expression and secretion of Yop proteins. Journal of Bacteriology, 191(13), 4259-4267
Open this publication in new window or tab >>Autoproteolysis of YscU of Yersinia pseudotuberculosis is important for regulation of expression and secretion of Yop proteins
2009 (English)In: Journal of Bacteriology, ISSN 0021-9193, E-ISSN 1098-5530, Vol. 191, no 13, p. 4259-4267Article in journal (Refereed) Published
Abstract [en]

YscU of Yersinia can be autoproteolysed to generate a 10-kDa C-terminal polypeptide designated YscU(CC). Autoproteolysis occurs at the conserved N downward arrowPTH motif of YscU. The specific in-cis-generated point mutants N263A and P264A were found to be defective in proteolysis. Both mutants expressed and secreted Yop proteins (Yops) in calcium-containing medium (+Ca(2+) conditions) and calcium-depleted medium (-Ca(2+) conditions). The level of Yop and LcrV secretion by the N263A mutant was about 20% that of the wild-type strain, but there was no significant difference in the ratio of the different secreted Yops, including LcrV. The N263A mutant secreted LcrQ regardless of the calcium concentration in the medium, corroborating the observation that Yops were expressed and secreted in Ca(2+)-containing medium by the mutant. YscF, the type III secretion system (T3SS) needle protein, was secreted at elevated levels by the mutant compared to the wild type when bacteria were grown under +Ca(2+) conditions. YscF secretion was induced in the mutant, as well as in the wild type, when the bacteria were incubated under -Ca(2+) conditions, although the mutant secreted smaller amounts of YscF. The N263A mutant was cytotoxic for HeLa cells, demonstrating that the T3SS-mediated delivery of effectors was functional. We suggest that YscU blocks Yop release and that autoproteolysis is required to relieve this block.

National Category
Medical and Health Sciences
Research subject
Medicine
Identifiers
urn:nbn:se:umu:diva-32213 (URN)10.1128/JB.01730-08 (DOI)19395493 (PubMedID)2-s2.0-67649400614 (Scopus ID)
Available from: 2010-03-03 Created: 2010-03-03 Last updated: 2023-03-24Bibliographically approved
Lavander, M., Ericsson, S. K., Bröms, J. E. & Forsberg, Å. (2007). Twin arginine translocation in Yersinia. Advances in Experimental Medicine and Biology, 603, 258-267
Open this publication in new window or tab >>Twin arginine translocation in Yersinia
2007 (English)In: Advances in Experimental Medicine and Biology, ISSN 0065-2598, E-ISSN 2214-8019, Vol. 603, p. 258-267Article in journal (Refereed) Published
Abstract [en]

Bacteria utilise Twin arginine translocation (Tat) to deliver folded proteins across the cytoplasmic membrane. Disruption of Tat typically results in pleiotropic effects on e.g. growth, stress resistance, bacterial membrane biogenesis, motility and cell morphology. Further, Tat is coupled to virulence in a range of pathogenic bacteria, including species of Pseudomonas, Legionella, Agrobacterium and Mycobacterium. We have investigated this, for Yersinia, previously unexplored system, and have shown that the Tat pathway is functional and absolutely required for virulence of Yersinia pseudotuberculosis. A range of putative Yersinia Tat substrates have been predicted in silico, which together with the Tat system itself may be interesting targets for future development of antimicrobial treatments. Here we present a brief review of bacterial Tat and discuss our results concerning this system in Yersinia.

Place, publisher, year, edition, pages
Springer, 2007
National Category
Bioinformatics and Systems Biology
Identifiers
urn:nbn:se:umu:diva-21150 (URN)10.1007/978-0-387-72124-8_23 (DOI)000250831600023 ()17966422 (PubMedID)
Available from: 2009-04-03 Created: 2009-04-03 Last updated: 2019-01-24Bibliographically approved
Lavander, M., Ericsson, S. K., Bröms, J. E. & Forsberg, Å. (2006). The Twin Arginine Translocation System is Essential for Virulence of Yersinia pseudotuberculosis. Infection and Immunity, 74(3), 1768-1776
Open this publication in new window or tab >>The Twin Arginine Translocation System is Essential for Virulence of Yersinia pseudotuberculosis
2006 (English)In: Infection and Immunity, ISSN 0019-9567, E-ISSN 1098-5522, Vol. 74, no 3, p. 1768-1776Article in journal (Refereed) Published
Abstract [en]

Yersinia species pathogenic to humans have been extensively characterized with respect to type III secretion and its essential role in virulence. This study concerns the twin arginine translocation (Tat) pathway utilized by gram-negative bacteria to secrete folded proteins across the bacterial inner membrane into the periplasmic compartment. We have shown that the Yersinia Tat system is functional and required for motility and contributes to acid resistance. A Yersinia pseudotuberculosis mutant strain with a disrupted Tat system (tatC) was, however, not affected in in vitro growth or more susceptible to high osmolarity, oxidative stress, or high temperature, nor was it impaired in type III secretion. Interestingly, the tatC mutant was severely attenuated via both the oral and intraperitoneal routes in the systemic mouse infection model and highly impaired in colonization of lymphoid organs like Peyer's patches and the spleen. Our work highlights that Tat secretion plays a key role in the virulence of Y. pseudotuberculosis.

Place, publisher, year, edition, pages
American Society for Microbiology, 2006
National Category
Bioinformatics and Systems Biology
Identifiers
urn:nbn:se:umu:diva-4775 (URN)10.1128/IAI.74.3.1768-1776.2006 (DOI)000235817500037 ()16495550 (PubMedID)2-s2.0-33644782616 (Scopus ID)
Available from: 2005-10-27 Created: 2005-10-27 Last updated: 2023-03-24Bibliographically approved
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