Mechanisms of recognition in dominant R gene mediated resistance.

One branch of plant innate immunity is mediated through what is traditionally known as race-specific or gene-for-gene resistance wherein the outcome of an attempted infection is determined by the genotypes of both the host and the pathogen. Dominant plant disease resistance (R) genes confer resistance to a variety of biotrophic pathogens, including viruses, encoding corresponding dominant avirulence (Avr) genes. R genes are among the most highly variable plant genes known, both within and between populations. Plant genomes encode hundreds of R genes that code for NB-LRR proteins, so named because they posses nucleotide-binding (NB) and leucine-rich repeat (LRR) domains. Many matching pairs of NB-LRR and Avr proteins have been identified as well as cellular proteins that mediate R/Avr interactions, and the molecular analysis of these interactions have led to the formulation of models of how products of R genes recognize pathogens. Data from multiple NB-LRR systems indicate that the LRR domains of NB-LRR proteins determine recognition specificity. However, recent evidence suggests that NB-LRR proteins have co-opted cellular recognition co-factors that mediate interactions between Avr proteins and the N-terminal domains of NB-LRR proteins.

Hydrographics and the timing of infectious salmon anemia outbreaks among Atlantic salmon (Salmo salar L.) farms in the Quoddy region of Maine, USA and New Brunswick, Canada.

Infectious salmon anemia (ISA) has caused severe morbidity and mortality in farmed Atlantic salmon in North America, Norway, Scotland and the Faroe Islands. The Quoddy region of Maine, United States of America (USA), and New Brunswick (NB), Canada is characterized by extensive tidal mixing and close proximity between farms. This region is also prone to recurrent appearances of ISA, though control measures limit disease spread and severity on infected farms. We conducted a retrospective longitudinal analysis of the apparent impact of hydrographics on the incidence and timing of ISA outbreaks on Atlantic salmon (Salmo salar L.) farms in the Quoddy region from May 2002 to August 2004. A time-series cross-sectional regression of 32 farms over 28 months demonstrated a limited, but statistically significant, spatio-temporal clustering of ISA outbreaks linked hydrographically. New outbreaks correlated temporally with those occurring on-site 1 and 3 months prior, and those occurring within one tidal-excursion upstream the same month. Other risk factors included holdover of previous year-class fish, wharf sharing, and possibly harvests of cages infected in previous months. Conclusions suggest that tidal dispersion does play a role in ISAV transmission in the Quoddy region. Dispersal of free virus and/or tidal distribution of lice or other hydrographically influenced vectors or fomites could all contribute to the spatio-temporal patterns described.

A comprehensive open reading frame phylogenetic analysis of isometric positive strand ssRNA plant viruses.

Rigorous large-scale whole genome comparisons are capable of providing more comprehensive and potentially more accurate descriptions of viral relationships, allowing for the effective validation and modification of current taxonomy. Using a set of 5 togaviruses as an outgroup, a comprehensive phylogeny for 115 isometric positive ssRNA plant viruses was generated based on the simultaneous comparison of over 480 ORFs found within completely sequenced genomes. With the exception of a diverse group of viruses representing the family Comoviridae, the single tree generated contained well supported branches corresponding to well established groups of viruses, including Bromoviridae, Umbravirus, Sobemovirus, and Tymoviridae. In addition, evidence for specific relationships between groups were also observed, specifically Tombusviridae + Umbravirus, and Luteoviridae + Sobemovirus. Various well established subgroups of viruses were also well resolved within the tree. In addition, some recent proposals involving the creation of new genera or the inclusion of newly described viruses into established genera were supported, while others were not. The evidence for frequent gene sharing and the potential consequences to viral taxonomy are discussed.

Different transcriptional properties of mSim-1 and mSim-2.

The mSim-1 and mSim-2 gene products are mammalian homologues of the Drosophila Sim gene. The dSim gene product transactivates through a DNA binding site known as the CNS midline enhancer (CME) element. We have investigated the transcriptional properties of mSIM-1 and mSIM-2 mediated through the CME element in concert with their dimerization partners, ARNT and ARNT-2. The mSIM-1/ARNT heterodimer transactivates reporter constructs via the ARNT carboxy-terminus. However, mSIM-2 quenches ARNT transactivation. We find that mSIM-2 competes with mSIM-1 for binding to ARNT, suggesting a possible antagonism between these transcription factors.

The Wilms' tumor suppressor gene (wt1) product represses different functional classes of transcriptional activation domains.

We have studied the ability of the wt1 tumor suppressor gene product to repress different classes of activation domains previously shown to stimulate the initiation and elongation steps of RNA polymerase II transcription in vivo. Repression assays revealed that WT1 represses all three classes of activation domains: Sp1 and CTF, which stimulate initiation (type I), human immunodeficiency virus type I Tat fused to a DNA-binding domain, which stimulates predominantly elongation (type IIA), and VP16, p53 and E2F1, which stimulate both initiation and elongation (type IIB). WT1 is capable of exerting its repression effect over a significant distance when positioned approximately 1700 bp from the core promoter. Deletion analysis of WT1 indicates that the responsible domain resides within the first 180 N-terminal amino acids of the protein. Nuclear run-ons analyzing the effects of WT1 on initiation of transcription demonstrate inhibition of this process. Our observations imply that WT1 can repress activators that stimulate initiation and/or elongation.

Regulation of renal EGF receptor expression is normal in Denys-Drash syndrome.

In patients with Denys-Drash syndrome, mutations of the Wilms' tumor suppressor gene are associated with nephroblastomas and developmental abnormalities of the genital tract and renal glomerulus. Normally, the Wilms' tumor gene product (WT1) is expressed at high levels in visceral glomerular epithelial cells (VGEC) of the emerging fetal glomerulus. We demonstrate that WT1 could normally serve to suppress EGF receptor expression in VGEC, since immunoreactive EGF receptor is strikingly absent compared to epithelial cells of the emerging proximal and distal tubule, which lack WT1. When HEK293 cells were co-transfected with plasmids containing EGFR enhancer/promoter elements linked to a CAT reporter and plasmids containing WT1 cDNA, EGFR enhancer/promoter activity was suppressed by all wild-type WT1 isoforms, but not by deletion mutants of WT1 lacking normal zinc-finger or N-terminal domains. Surprisingly, plasmids expressing a Denys-Drash WT1 mutant (R394W) retained the ability to suppress EGFR promoter activity in this system. Furthermore, we found that immunoreactive EGFR was appropriately undetectable in glomeruli from a three-year-old girl with Denys-Drash syndrome and in sections of her Wilm's tumor. These data suggest that faulty suppression of EGFR cannot account for the abnormalities of glomerulogenesis seen in Denys-Drash patients.

The murine Sim-2 gene product inhibits transcription by active repression and functional interference.

The Drosophila single-minded (Dsim) gene encodes a master regulatory protein involved in cell fate determination during midline development. This protein is a member of a rapidly expanding family of gene products possessing basic helix-loop-helix (bHLH) and hydrophobic PAS (designated a conserved region among PER, ARNT [aryl hydrocarbon receptor nuclear translocator] and SIM) protein association domains. Members of this family function as central transcriptional regulators in cellular differentiation and in the response to environmental stimuli such as xenobiotics and hypoxia. We have previously identified a murine member of this family, called mSim-2, showing sequence homology to the bHLH and PAS domains of Dsim. Immunoprecipitation experiments with recombinant proteins indicate that mSIM-2 associates with the arnt gene product. In the present work, by using fine-structure mapping we found that the HLH and PAS motifs of both proteins are required for optimal association. Forced expression of GAL4/mSIM-2 fusion constructs in mammalian cells demonstrated the presence of two separable repression domains within the carboxy terminus of mSIM-2. We found that mSIM-2 is capable of repressing ARNT-mediated transcriptional activation in a mammalian two-hybrid system. This effect (i) is dependent on the ability of mSIM-2 and ARNT to heterodimerize, (ii) is dependent on the presence of the mSIM-2 carboxy-terminal repression domain, and (iii) is not specific to the ARNT activation domain. These results suggest that mSIM-2 repression activity can dominantly override the activation potential of adjacent transcription factors. We also demonstrated that mSIM-2 can functionally interfere with hypoxia-inducible factor 1alpha (HIF-1alpha)/ARNT transcription complexes, providing a second mechanism by which mSIM-2 may inhibit transcription.

Identification of nuclear localization signals within the zinc fingers of the WT1 tumor suppressor gene product.

WT1 encodes a zinc finger protein with a key role in urogenital development that is inactivated in a subset of Wilms' tumors. This tumor suppressor gene product contains an amino-terminal dimerization domain required for trans-inhibition of wild-type WT1 activity by mutants defective for DNA binding. In the course of characterizing truncation mutants of WT1, we noted that the WT1 zinc fingers contain two functionally independent targeting signals required for nuclear localization of the protein. These novel signals lie within zinc fingers I and within zinc fingers II and III. We demonstrate that nuclear targeting of the WT1 homodimerization domain functionally antagonizes activity of the wild-type protein activity.

Characterization of msim, a murine homologue of the Drosophila sim transcription factor.

Mutations in the Drosophila single-minded (sim) gene result in loss of precursor cells that give rise to midline cells of the embryonic central nervous system. During the course of an exon-trapping strategy aimed at identifying transcripts that contribute to the etiology and pathophysiology of Down syndrome, we identified a human exon from the Down syndrome critical region showing significant homology to the Drosophila sim gene. Using a cross-hybridization approach, we have isolated a murine homolog of the Drosophila sim gene, which we designated msim. Nucleotide and predicted amino acid sequence analyses of msim cDNA clones indicate that this gene encodes a member of the basic-helix-loop-helix class of transcription factors. The murine and Drosophila proteins share 88% residues within the basic-helix-loop-helix domain, with an overall homology of 92%. In addition, the N-terminal domain of MSIM contains two PAS dimerization motifs also featured in the Drosophila sim gene product, as well as a small number of other transcription factors. Northern blot analysis of adult murine tissues revealed that the msim gene produces a single mRNA species of approximately 4 kb expressed in a small number of tissues, with the highest levels in the kidneys and lower levels present in skeletal muscle, lung, testis, brain, and heart. In situ hybridization experiments demonstrate that msim is also expressed in early fetal development in the central nervous system and in cartilage primordia. The characteristics of the msim gene are consistent with its putative function as a transcriptional regulator.

Geriatric rehabilitation: nursing interventions and outcomes focusing on urinary function and knowledge of medications.

This article describes the nursing interventions that focus on urinary function and medication knowledge of older adults in a geriatric rehabilitation program. The authors review the interventions needed to achieve bladder retraining and self-administration of medication and consider what predicts urinary function and knowledge of medication administration in older adults. The descriptive study included 251 consecutive patients who were admitted into a geriatric rehabilitation unit. An evaluation of urinary function was done on admission and based on patient need to (a) maintain urinary function, (b) decrease urinary incontinence, and (c) resolve urinary retention. All patients received routine medication teaching from admission to discharge during each daytime administration of medication. From admission to discharge, there was a decrease in incidences of urinary incontinence and urinary retention and an increase in knowledge of medication regimens. Although the study was descriptive, the findings suggested that nursing interventions in geriatric rehabilitation may decrease urinary incontinence and retention and improve knowledge of medication regimens in this population.

Antagonism of WT1 activity by protein self-association.

Germline loss-of-function mutations at the Wilms tumor (WT) suppressor locus WT1 are associated with a predisposition to WTs and mild genital system anomalies. In contrast, germ-line missense mutations within the WT1 gene encoding the DNA-binding domain often yield a more severe phenotype consisting of WT, sexual ambiguity, and renal nephropathy. In this report, we demonstrate that the products of mutant alleles that impair DNA recognition can antagonize WT1-mediated transcriptional repression. We demonstrate that WT1 can self-associate in vitro and in vivo and that the responsible domain maps to the amino-terminal region of the protein. Oligomers of full-length protein form less efficiently or produce less stable complexes than oligomers between truncated polypeptides and full-length protein. Our data suggest a molecular mechanism to explain how WT1 mutations may act in deregulating cellular proliferation and differentiation.

Somatization disorder (Briquet's syndrome) in a male veteran.

The authors present here a case of somatization disorder in a heterosexual male veteran. The age at onset of 7 years is the earliest yet reported in a male. Extensive medical records provide a unique opportunity to follow the course of a relatively severe case of this condition and demonstrate that it fulfills the most rigorous diagnostic criteria that have been formulated. Yet, the case of Orville N is more than a lengthy list of medically unexplained symptoms, showing as it does a number of other features traditionally associated with the broad category of hysterical psychopathology.