| 1. |
- Kollmus, H., et al.
(författare)
-
Energy scaling of the ion-induced desorption yield for perpendicular collisions of Ar and U with stainless steel in the energy range of 5 and 100 MeV/u
- 2009
-
Ingår i: Journal of Vacuum Science & Technology. A. Vacuum, Surfaces, and Films. - : American Vacuum Society. - 0734-2101 .- 1520-8559. ; 27:2, s. 245-247
-
Tidskriftsartikel (refereegranskat)abstract
- For the GSI future project Facility for Antiproton and Ion Research a beam intensity of 10(12)U(28+)ions/s is planned to be extracted from the GSI heavy ion synchrotron SIS18. Measurements performed in 2001 showed that the beam lifetime of the ions in the synchrotron is decreasing with increasing number of injected particles due to vacuum instabilities caused by ion-induced desorption. The injection energy for the SIS18 is about 10 MeV/u and U28+ ions are accelerated to 200 MeV/u limited by the magnetic rigidity for the low charge state. The aim of this work was to measure the desorption yield as a function of the impact energy from injection to extraction of SIS18 at GSI. Low energy yields at 5.0, 9.7, and 17.7 MeV/u were measured at the Cyclotron of The Svedberg Laboratory in Uppsala. High energy yields at 40, 80, and 100 MeV/u were measured at SIS18 of GSI in a different setup. It was found that the desorption yield scales with the electronic energy loss (dE/dx)(el)(n), with n between 2 and 3, decreasing for increasing impact energy above the Bragg maximum.
|
|
| 2. |
- Bröms, Jeanette E, et al.
(författare)
-
Mapping of a YscY binding domain within the LcrH chaperone that is required for regulation of Yersinia type III secretion
- 2005
-
Ingår i: Journal of Bacteriology. - : American Society for Microbiology. - 0021-9193 .- 1098-5530. ; 187:22, s. 7738-7752
-
Tidskriftsartikel (refereegranskat)abstract
- Type III secretion systems are used by many animal and plant interacting bacteria to colonize their host. These systems are often composed of at least 40 genes, making their temporal and spatial regulation very complex. Some type III chaperones of the translocator class are important regulatory molecules, such as the LcrH chaperone of Yersinia pseudotuberculosis. In contrast, the highly homologous PcrH chaperone has no regulatory effect in native Pseudomonas aeruginosa or when produced in Yersinia. In this study, we used LcrH-PcrH chaperone hybrids to identify a discrete region in the N terminus of LcrH that is necessary for YscY binding and regulatory control of the Yersinia type III secretion machinery. PcrH was unable to bind YscY and the homologue Pcr4 of P. aeruginosa. YscY and Pcr4 were both essential for type III secretion and reciprocally bound to both substrates YscX of Yersinia and Pcr3 of P. aeruginosa. Still, Pcr4 was unable to complement a DeltayscY null mutant defective for type III secretion and yop-regulatory control in Yersinia, despite the ability of YscY to function in P. aeruginosa. Taken together, we conclude that the cross-talk between the LcrH and YscY components represents a strategic regulatory pathway specific to Yersinia type III secretion.
|
|
| 3. |
- Hedlund, E., et al.
(författare)
-
A new test stand for heavy ion induced gas desorption measurements at TSL
- 2008
-
Ingår i: Nuclear Instruments and Methods in Physics Research Section A. - : Elsevier BV. - 0168-9002 .- 1872-9576. ; 586:3, s. 377-381
-
Tidskriftsartikel (refereegranskat)abstract
- In several experiments at CERN, GSI and BNL it has been found that the lifetime of highly energetic heavy ions in synchrotrons decreases with increasing number of injected ions. This phenomenon occurs due to the collisions of beam ions and residual gas molecules leading to the change of charge of the ions and their loss on the vacuum chamber walls, which in turn cause ion-induced gas desorption and further pressure increase. To gain a deeper understanding of the ion-induced desorption process in the energy range 5-45 MeV/u, a dedicated test stand was built at the end of the K beamline at The Svedberg Laboratory (TSL) in Uppsala, Sweden. The energy range was chosen due to the fact that the injection energy of the heavy ion synchrotron SIS18 at GSI will be 10 MeV/u, and that there are insufficient data in this energy range. A Test Particle Monte-Carlo model of the experimental set-up was build-up, run and analysed for different sample configurations. An important result is that for the same sample material the desorption yield from a flat sample causes a 1.58 times larger pressure increase than that of a tubular sample. A detailed explanation of the set-up is presented.
|
|
| 4. |
- Hedlund, E., et al.
(författare)
-
Ar ion induced desorption yields at the energies 5-17.7 MeV/u
- 2009
-
Ingår i: Nuclear Instruments and Methods in Physics Research Section A. - : Elsevier BV. - 0168-9002 .- 1872-9576. ; 599:1, s. 1-8
-
Tidskriftsartikel (refereegranskat)abstract
- Particle accelerators have, during Operation with heavy ion beams, shown a significant pressure rise when the intensity of the beam is increased. This pressure rise is due to ion induced desorption, which is the result of beam ions colliding with residual gas atoms in the beam pipe, where they undergo charge exchange. This causes them to hit the vacuum chamber after the next dipole magnet and gas to be released. For the upgrade of the SIS18 synchrotron at GSI the intensity has to be a few orders of magnitude higher than it is today at the injection energy of 10 MeV/u. The aim of this experiment is to measure desorption yields, n, (released Molecules per incident ion) from materials commonly used in accelerators: 316LN stainless steel, Cu, etched Cu, gold coated Cu and Ta, using an Ar beam at impact energies in the range of 5-17.7 MeV/u for perpendicular incidence. The measured initial desorption yields vary for the same material from sample to sample: up to 4.5 times for stainless steel and Lip to 3 times for etched Cu. Therefore more samples should be Studied to have better statistics. Beam conditioning at lower energy does not significantly reduce the desorption yield at higher energy. There is a significant difference Of Lip to a few times in desorption yield between flat and tubular samples. The desorption yield from a Cu sample at grazing incident angle of 125 mrad was an order of magnitude larger than at normal incident angle. It Was found that the total number of positively and negatively charged secondary particles, emitted from the Surface bombarded with heavy ions, does not exceed -40 secondary particles per impact heavy ion. The current of negatively charged particles was about 2.3 times larger than the current for positively charged particles. The impact from secondary particles on dynamic gas pressure was not possible to investigate.
|
|
| 5. |
|
|
| 6. |
- Bröms, Jeanette E, et al.
(författare)
-
Tetratricopeptide repeats are essential for PcrH chaperone function in Pseudomonas aeruginosa type III secretion.
- 2006
-
Ingår i: FEMS Microbiol Lett. - 0378-1097. ; 256:1, s. 57-66
-
Tidskriftsartikel (refereegranskat)abstract
- The type III secretion system (T3SS) is a specialized apparatus evolved by Gram-negative bacteria to deliver effector proteins into host cells, thus facilitating the establishment of an infection. Effector translocation across the target cell plasma membrane is believed to occur via pores formed by at least two secreted translocator proteins, the functions of which are dependent upon customized class II T3SS chaperones. Recently, three internal tetratricopeptide repeats (TPRs) were identified in this class of chaperones. Here, defined mutagenesis of the class II chaperone PcrH of Pseudomonas aeruginosa revealed these TPRs to be essential for chaperone activity towards the translocator proteins PopB and PopD and subsequently for the translocation of exoenzymes into host cells.
|
|
| 7. |
- Carlsson, Katrin E, et al.
(författare)
-
Extracytoplasmic-stress-responsive pathways modulate type III secretion in Yersinia pseudotuberculosis.
- 2007
-
Ingår i: Infect Immun. - 0019-9567. ; 75:8, s. 3913-24
-
Tidskriftsartikel (refereegranskat)abstract
- Three signal transduction pathways, the two-component systems CpxRA and BaeSR and the alternative sigma factor sigma(E), respond to extracytoplasmic stress that facilitates bacterial adaptation to changing environments. At least the CpxRA and sigma(E) pathways control the production of protein-folding and degradation factors that counter the effects of protein misfolding in the periplasm. This function also influences the biogenesis of multicomponent extracellular appendages that span the bacterial envelope, such as various forms of pili. Herein, we investigated whether any of these regulatory pathways in the enteropathogen Yersinia pseudotuberculosis affect the functionality of the Ysc-Yop type III secretion system. This is a multicomponent molecular syringe spanning the bacterial envelope used to inject effector proteins directly into eukaryotic cells. Disruption of individual components revealed that the Cpx and sigma(E) pathways are important for Y. pseudotuberculosis type III secretion of Yops (Yersinia outer proteins). In particular, a loss of CpxA, a sensor kinase, reduced levels of structural Ysc (Yersinia secretion) components in bacterial membranes, suggesting that these mutant bacteria are less able to assemble a functional secretion apparatus. Moreover, these bacteria were no longer capable of localizing Yops into the eukaryotic cell interior. In addition, a cpxA lcrQ double mutant engineered to overproduce and secrete Yops was still impaired in intoxicating cells. Thus, the Cpx pathway might mediate multiple influences on bacterium-target cell contact that modulate Yersinia type III secretion-dependent host cell cytotoxicity.
|
|
| 8. |
- Carlsson, Katrin E, et al.
(författare)
-
Influence of the Cpx extracytoplasmic-stress-responsive pathway on Yersinia sp.-eukaryotic cell contact.
- 2007
-
Ingår i: Infect Immun. - 0019-9567. ; 75:9, s. 4386-99
-
Tidskriftsartikel (refereegranskat)abstract
- The extracytoplasmic-stress-responsive CpxRA two-component signal transduction pathway allows bacteria to adapt to growth in extreme environments. It controls the production of periplasmic protein folding and degradation factors, which aids in the biogenesis of multicomponent virulence determinants that span the bacterial envelope. This is true of the Yersinia pseudotuberculosis Ysc-Yop type III secretion system. However, despite using a second-site suppressor mutation to restore Yop effector secretion by yersiniae defective in the CpxA sensor kinase, these bacteria poorly translocated Yops into target eukaryotic cells. Investigation of this phenotype herein revealed that the expression of genes which encode several surface-located adhesins is also influenced by the Cpx pathway. In particular, the expression and surface localization of invasin, an adhesin that engages beta1-integrins on the eukaryotic cell surface, are severely restricted by the removal of CpxA. This reduces bacterial association with eukaryotic cells, which could be suppressed by the ectopic production of CpxA, invasin, or RovA, a positive activator of inv expression. In turn, these infected eukaryotic cells then became susceptible to intoxication by translocated Yop effectors. In contrast, bacteria harboring an in-frame deletion of cpxR, which encodes the cognate response regulator, displayed an enhanced ability to interact with cell monolayers, as well as elevated inv and rovA transcription. This phenotype could be drastically suppressed by providing a wild-type copy of cpxR in trans. We propose a mechanism of inv regulation influenced by the direct negative effects of phosphorylated CpxR on inv and rovA transcription. In this fashion, sensing of extracytoplasmic stress by CpxAR contributes to productive Yersinia sp.-eukaryotic cell interactions.
|
|
| 9. |
- Carreras-Puigvert, Jordi, et al.
(författare)
-
A comprehensive structural, biochemical and biological profiling of the human NUDIX hydrolase family
- 2017
-
Ingår i: Nature Communications. - : Nature Publishing Group. - 2041-1723. ; 8:1
-
Tidskriftsartikel (refereegranskat)abstract
- The NUDIX enzymes are involved in cellular metabolism and homeostasis, as well as mRNA processing. Although highly conserved throughout all organisms, their biological roles and biochemical redundancies remain largely unclear. To address this, we globally resolve their individual properties and inter-relationships. We purify 18 of the human NUDIX proteins and screen 52 substrates, providing a substrate redundancy map. Using crystal structures, we generate sequence alignment analyses revealing four major structural classes. To a certain extent, their substrate preference redundancies correlate with structural classes, thus linking structure and activity relationships. To elucidate interdependence among the NUDIX hydrolases, we pairwise deplete them generating an epistatic interaction map, evaluate cell cycle perturbations upon knockdown in normal and cancer cells, and analyse their protein and mRNA expression in normal and cancer tissues. Using a novel FUSION algorithm, we integrate all data creating a comprehensive NUDIX enzyme profile map, which will prove fundamental to understanding their biological functionality.
|
|
| 10. |
- Costa, Tiago R. D., et al.
(författare)
-
Influence of the LcrH chaperone on type III secretion system regulation in Yersinia pseudotuberculosis
-
Annan publikation (övrigt vetenskapligt/konstnärligt)abstract
- Human pathogenic Yersiniae share a common virulence plasmid that encodes for the Ysc-Yop type III secretion system (T3SS). Control of yop expression involves several pathways in which their cross-talk is not completely understood. LcrF, an AraC-like transcriptional activator, is required for temperature-dependent yop-transcription. In contrast, a repressive effect of the T3S chaperone LcrH and the cognate translocator substrate YopD occurs through binding to yop mRNA and inhibiting translation; a process that is also thought to involve LcrQ. Several homologous members of the LcrH family of translocator-class of T3S chaperones can act as a cofactor to amplify the activity of transcriptional activators analogous to LcrF. However, we show here in Y. pseudotuberculosis that LcrH does not induce LcrF-dependent transcription of target genes. Moreover, a full length DlcrH null mutant in which YopB and YopD are rapidly degraded is totally de-repressed for Yop synthesis even though the anti-activator LcrQ is forced to accumulate in the cytoplasm through rendering the Ysc-Yop T3SS non-functional or ectopically producing LcrQ in trans. Typically, this mutant cannot grow at 37°C. Thus, in all respects, the DlcrH null mutant mirrors the regulatory defects established for Yersinia lacking the translocator and anti-activator YopD. On the other hand, Y. pseudotuberculosis producing the LcrHE30G point mutant that is defective for YscY chaperone binding exhibits a mild regulatory defect that permits some growth at 37°C, but is blind to the cytoplasmic accumulation of LcrQ. Critically however, this mutant still responds to repression caused by YopD accumulation, which is stably produced and efficiently secreted by this strain. Thus, our work with LcrHE30G indicates an additional regulatory function of this versatile T3S chaperone that is independent of the LcrF transcription factor and the YopD anti-activator.
|
|
