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A Forsberg

Publications and source records attributed to A Forsberg.

At least 37 records · Page 2Linked to original sources
PubMed1995

Bacterial evasion of host immune defense: Yersinia enterocolitica encodes a suppressor for tumor necrosis factor alpha expression.

The ability of the enteropathogenic Yersinia enterocolitica to survive and proliferate in host tissue depends on a 70-kb plasmid known to encode a number of released Yersinia outer proteins that act as virulence factors by inducing cytotoxicity and inhibiting phagocytosis. This study demonstrates that one of the Yersinia outer proteins, the 41-kDa YopB, suppresses the production of tumor necrosis factor alpha (TNF-alpha), a macrophage-derived cytokine with central roles in the regulation of immune and inflammatory responses to infection. This conclusion is based on several lines of evidence. First, in macrophage cultures, suppression of TNF-alpha mRNA expression was induced by culture supernatant (CS+) of plasmid-bearing yersiniae, the effect which was blocked by anti-YopB antiserum. Second, suppression of TNF-alpha production, but not of interleukin-1 (IL-1) and IL-6, was induced by purified YopB. Third, in Yersinia-infected mice, no increase in TNF-alpha mRNA expression was observed in Peyer's patches, the primary site of bacterial invasion, compared with IL-1 (alpha and beta) mRNA. Finally, administration of anti-YopB antiserum to mice prior to infection with Y. enterocolitica increased TNF activity levels in Peyer's patches and coincided with a reduction in bacterial growth. The results thus provide direct evidence for a secreted eubacterial virulence factor that mediates selective suppression of TNF-alpha production. Although suppression of this single cytokine response is probably not sufficient to facilitate survival of the infecting organisms, the results suggest that suppression of TNF-alpha production by YopB significantly contributes to the evasion of Y. enterocolitica from antibacterial host defense.

Animals↗
PubMed1995

Virulence plasmid-encoded YopK is essential for Yersinia pseudotuberculosis to cause systemic infection in mice.

The virulence plasmid common to pathogenic Yersinia species encodes a number of secreted proteins denoted Yops (Yersinia outer proteins). Here, we identify and characterize a novel plasmid-encoded virulence determinant of Yersinia pseudotuberculosis, YopK. The yopK gene was found to be conserved among the three pathogenic Yersinia species and to be homologous to the previously described yopQ and yopK genes of Y. enterocolitica and Y. pestis, respectively. Similar to the other Yops, YopK expression and secretion were shown to be regulated by temperature and by the extracellular Ca2+ concentration; thus, yopK is part of the yop regulon. In addition, YopK secretion was mediated by the specific Yop secretion system. In Y. pseudotuberculosis, YopK was shown neither to have a role in this bacterium's ability to resist phagocytosis by macrophages nor to cause cytotoxicity in HeLa cells. YopK was, however, shown to be required for the bacterium to cause a systemic infection in both intraperitoneally and orally infected mice. Characterization of the infection kinetics showed that, similarly to the wild-type strain, the yopK mutant strain colonized and persisted in the Peyer's patches of orally infected mice. A yopE mutant which is impaired in cytotoxicity and in antiphagocytosis was, however, found to be rapidly cleared from these lymphoid organs. Neither the yopK nor the yopE mutant strain could overcome the primary host defense and reach the spleen. This finding implies that YopK acts at a different level during the infections process than the antiphagocytic YopE cytotoxin does.

Animals↗
PubMed1993

A secreted protein kinase of Yersinia pseudotuberculosis is an indispensable virulence determinant.

Phosphorylation of proteins catalysed by protein kinases is associated with central functions in growth and proliferation of the eukaryotic cell, and kinases are particularly important in the signal transduction pathways. Enterobacterial protein kinases are structurally and functionally different from eukaryotic protein kinases, and no prokaryotic kinase has so far been described implicating a direct role for this activity in virulence. Virulent Yersinia possess a common virulence plasmid that encodes a number of secreted proteins (Yops), of which YopH has protein-tyrosine phosphatase activity with a key function in the block of phagocytosis by the pathogen. Here we report that the virulence plasmid of Yersinia pseudotuberculosis encodes a secreted protein kinase (YpkA) with extensive homology to eukaryotic Ser/Thr protein kinases. Specific mutants of ypkA resulted in avirulent strains. Thus, YpkA is, to our knowledge, the first reported prokaryotic secreted protein kinase involved in pathogenicity, presumably by interfering with the signal transduction pathways of the target cell.

Amino Acid Sequence↗
PubMed1993

In vivo expression of virulence genes of Yersinia pseudotuberculosis.

The common virulence plasmid of pathogenic Yersinia species encodes a number of secreted proteins denoted Yops. YopE and YopH are indispensable for virulence and act in concert to mediate resistance to phagocytosis. In vitro all yop genes are coordinately regulated, with maximal expression and secretion at 37 degrees C in medium lacking Ca2+. In order to facilitate studies of in vivo expression of yopE and yopH we constructed operon fusions between these two genes and a promoterless luxAB operon from Vibrio harveyi in Y. pseudotuberculosis. Groups of 10 mice were infected with each fusion strain, and colonization and luciferase expression in Peyer's patches and spleen were followed by sacrificing two mice from each group daily. Peyer's patches were found to be colonized from the first day of infection, whereas the spleen was not colonized until the third day of infection. Luciferase expression could easily be monitored if the sample contained > or = 10(4) bacteria. Expression levels tended to be highest at the early stages of colonization for the respective organ (day 1 for Peyer's patches and day 3 or 4 for spleen). When in vivo expression was compared with in vitro expression, it was found that during infection the expression levels were as high or even higher than the in vitro expression observed for strains grown at 37 degrees C in Ca(2+)-depleted media.

Bacterial Outer Membrane Proteins↗
PubMed1992

A novel protein, LcrQ, involved in the low-calcium response of Yersinia pseudotuberculosis shows extensive homology to YopH.

The plasmid-encoded yop genes of pathogenic yersiniae are regulated by the environmental stimuli calcium and temperature. A novel protein, LcrQ, which exhibits a key function in the negative calcium-controlled pathway, was identified. DNA sequence analysis revealed that LcrQ has a molecular mass of 12,412 daltons and its isoelectric point is 6.51. Overexpression of LcrQ in trans in wild-type Yersinia pseudotuberculosis YPIII(pIB102) changed the phenotype from calcium dependence to calcium independence and inhibited Yop expression. LcrQ is expressed from a monocistronic operon. Trans overexpression of LcrQ in yopN and lcrH mutants affected the phenotype of the yopN mutant (temperature sensitive to calcium independence) but not that of the lcrH mutant (temperature sensitive), suggesting that LcrQ acts between YopN and LcrH in the calcium-regulated pathway. An lcrQ mutant was found to be temperature sensitive for growth and showed derepressed Yop expression at 37 degrees C in the presence of calcium in the growth medium. During these culture conditions, the lcrQ mutant secreted only LcrV and YopD into the culture supernatant. Removal of Ca2+ from the growth medium resulted in a Yop expression pattern of the mutant that was identical to that of the wild-type strain. The LcrQ protein was recovered from the culture supernatant. LcrQ shows 42% identity to the first 128 amino acids of the YopH virulence protein.

Amino Acid Sequence↗
PubMed1992

Influence of body mass on cross-country ski racing performance.

The influence of body mass on cross-country skiing performance was evaluated by (i) a theoretical analysis of the relationship between body mass (M) and on one hand the maximal aerobic power and on the other the power expense of skiing, (ii) timing skiers in varied terrain during ski racing, (iii) comparing racing success between light and heavy skiers, and (iv) relating maximal aerobic power of skiers and power expense of skiing to body mass. The theoretical analysis indicated that the maximal aerobic power scaled with M2/3, while the mass exponent for the power expenses ranged from 1/3 to 1. The net result was that heavy skiers appear to be favored in all types of terrain but the steep uphills. During ski racing, heavy skiers displayed a tendency to be faster in all types of terrain, although no statistically significant effect was found. The racing success tended to be greater in heavy skiers among males but not among females. During skiing, maximal aerobic power and the power expense of skiing scaled with M0.7 and M0.4, respectively, indicating an advantage for the heavy skier. Altogether, these observations indicate that heavy skiers are favored, but also that other factors are more important than body mass for cross-country ski racing performance.

Body Height↗
PubMed1991

The surface-located YopN protein is involved in calcium signal transduction in Yersinia pseudotuberculosis.

The low-calcium response (lcr) is strongly conserved among the pathogenic Yersinia species and is observed when the pathogen is grown at 37 degrees C in Ca(2+)-depleted medium. This response is characterized by a general metabolic downshift and by a specific induction of virulence-plasmid-encoded yop genes. Regulation of yop expression is exerted at transcriptional level by a temperature-regulated activator and by Ca(2+)-regulated negative elements. The yopN gene was shown to encode a protein (formerly also designated Yop4b) which is surface-located when Yersinia is grown at 37 degrees C. yopN was found to be part of an operon that is induced during the low-calcium response. Insertional inactivation of the yopN gene resulted in derepressed transcription of yop genes. A hybrid plasmid containing the yopN gene under the control of the tac promoter fully restored the wild-type phenotype of the yopN mutant. Thus the surface-located YopN somehow senses the calcium concentration and transmits a signal to shut off yop transcription when the calcium concentration is high.

Amino Acid Sequence↗
PubMed1991

Intracellular targeting of the Yersinia YopE cytotoxin in mammalian cells induces actin microfilament disruption.

Pathogenic Yersinia spp., including the etiological agent of plague, Y. pestis, all carry a common plasmid that encodes a number of essential virulence determinants, the Yop proteins. One of these, YopE, has been shown to be involved in the obstruction of the primary host defense by a molecular mechanism leading to inhibition of phagocytosis (R. Rosqvist, A. Forsberg, M. Rimpiläinen, T. Bergman, and H. Wolf-Watz, Mol. Microbiol. 4:657-667, 1990). Although the Yop proteins are secreted into the culture supernatant in vast amounts, in vitro studies of the function of the Yop proteins have so far been unsuccessful. We show that isolated Yop proteins indeed can cause cytotoxic effects in vitro if the proteins are introduced intracellularly into the eukaryotic cell. Isolated Yop proteins of Yersinia pseudotuberculosis were found to disrupt the microfilament structure when microinjected intracellularly into the host cell. In particular, YopE was demonstrated to be directly involved in the cytotoxic action, whereas YopD seems to have a critical role in translocating the YopE protein through the host cell membrane. These results elucidate the requirement for at least some of the Yop proteins to leave the pathogen during infection.

Actin Cytoskeleton↗
PubMed1991

Analysis of the V antigen lcrGVH-yopBD operon of Yersinia pseudotuberculosis: evidence for a regulatory role of LcrH and LcrV.

Virulent Yersinia species possess a common plasmid that encodes essential virulence determinants (Yops) which are regulated by the extracellular stimuli Ca2+ and temperature. The V antigen operon was recently shown to be involved in the Ca2(+)-regulated negative pathway (A. Forsberg and H. Wolf-Watz, Mol. Microbiol. 2:121-133, 1988). We show here that the V antigen-containing operon of Yersinia pseudotuberculosis is a polycistronic operon having the gene order lcrGVH-yopBD. DNA sequencing analysis of lcrGVH revealed a high homology to the corresponding genes of Yersinia pestis. LcrG was conserved and LcrH showed only one amino acid difference, while LcrV showed only 96.6% identity. The amino acid substitutions of LcrV occurred in the central domain of the protein, while the two ends of the protein were conserved. Northern (RNA) blotting experiments showed that the operon is regulated at the transcriptional level by the extracellular stimuli temperature and calcium. One 4.6-kb transcriptional product of the operon was identified. This mRNA is rapidly processed at its 5' end, resulting in different mRNA species of variable stability. By genetic analysis, the lcrV and lcrH gene products were found to be regulatory proteins having important roles in the Ca2(+)-controlled regulation of Yop expression. The activity of LcrH is modulated by a gene product of the operon that inhibits the negative action of LcrH on yop transcription in the absence of Ca2+.

Amino Acid Sequence↗
PubMed1991

The relationship between body mass and oxygen uptake during running in humans.

Oxygen uptake during treadmill running was measured at submaximal and maximal intensities in six different groups of endurance athletes (N = 134) and in seven endurance-trained men. The relationship between body mass (M) and oxygen uptake (VO2) was evaluated by deriving the exponent b in the equation VO2 = a.Mb. Thus, if b = 1, the oxygen uptake increases in proportion to body mass and oxygen uptake per kg is independent of body mass; if b less than 1, than the oxygen uptake per kg is inversely related to body mass. The exponent b was found to be less than unity for all groups for both submaximal (b = 0.76, s = 0.06) and maximal oxygen uptake (b = 0.71, s = 0.05). These results indicate that neither submaximal nor maximal oxygen uptake increases in proportion to body mass during running. The relationship between submaximal oxygen uptake and body mass observed in this study may explain why the oxygen uptake per kg of body mass has been found to be higher for children than for adults.

Adolescent↗
PubMed1990

The cytotoxic protein YopE of Yersinia obstructs the primary host defence.

It has previously been shown that the plasmid-encoded YopE protein of Yersinia pseudotuberculosis is a virulence determinant. In this study, HeLa cells, macrophages and mice were used as different model systems to determine the actual role of YopE in the virulence process. The YopE protein mediates a cytotoxic response on a confluent layer of HeLa cells. A prerequisite of this activity is that the pathogen binds to the cell surface. YopE also induces a cytotoxic response on mouse macrophages where it influences the ability of the pathogen to resist phagocytosis. Bacterial mutants defective in their ability to express YopE are avirulent after oral or intraperitoneal infection but virulent following intravenous injection. On the basis of these results, we propose a role for YopE in the virulence process of Yersinia.

Animals↗