Inhibition of Nucleolar Proteins in <italic>Caenorhabditis Elegans</Italic> Confers Enhanced Resistance to <italic>Salmonella Enterica</Italic> through a P53/<Italic>cep-1</Italic>-Dependent Mechanism

dc.contributor.advisor

Aballay, Alejandro

dc.contributor.advisor

Marchuk, Douglas

dc.contributor.author

Fuhrman, Laura Elizabeth

dc.date.accessioned

2009-05-01T18:32:48Z

dc.date.available

2011-07-26T04:30:04Z

dc.date.issued

2009

dc.department

Genetics and Genomics

dc.description.abstract

The relatively simple innate immune system of Caenorhabditis elegans and the number of traits that facilitate genetic and genomic analysis using this organism have nurtured rapid advances into the understanding of C. elegans innate immunity during the last few years. However, traditional methods of isolating and mapping C. elegans mutants exhibiting aberrant immune responses to pathogen infection are often labor intensive and time consuming. Therefore, a simple and rapid means of isolating and mapping C. elegans immune mutants will increase the number of mutants that can be studied. Salmonella enterica, as well as other bacterial pathogens, has been described to cause a significant distension of the C. elegans intestinal lumen, which correlates with death of the nematode. C. elegans mutants which exhibit a weakened immune response would therefore be expected to develop intestinal distension at an earlier time point than wild type. Likewise, mutants which exhibit an enhanced immune response would be expected to develop intestinal distension at a later time point than wild type. Taking advantage of this correlation, we designed a novel approach to isolating C. elegans mutants which exhibit aberrant immune responses to the bacterial pathogen, S. enterica. Furthermore, we validated and optimized the use of Amplifluor®, a high-throughput genotyping system, for use in C. elegans single nucleotide polymorphism (SNP) mapping.

To date, the only known negative regulators of innate immunity in C. elegans are dependent on the FOXO transcription factor, DAF-16 and regulate lifespan in addition to immunity. Therefore, we focused our efforts on identifying additional negative regulators of innate immunity by screening for mutants which display a reduced accumulation of S. enterica at a time point when wild-type nematodes are packed with bacteria. In a genetic screen for C. elegans mutants which display reduced accumulation of S. enterica/GFP, we identified a mutation in nol-6, a nucleolar protein containing a nucleolar RNA-associated protein (Nrap) domain which is conserved across eukaryotic organisms. nol-6 is implicated in ribosomal RNA (rRNA) processing during the early stages of ribosome biogenesis. We show that knockdown of nol-6 as well as other nucleolar genes leads to a reduction of pathogen accumulation and enhanced resistance to killing by pathogen. In addition, we demonstrate that enhanced resistance is dependent on p53/cep-1. Furthermore, microarray analysis shows a significant enrichment of upregulated genes that have previously been shown to be dependent on p53/cep-1 for induction following ultraviolet radiation. These results represent the first evidence that C. elegans innate immunity is regulated by the nucleolus through a p53/cep-1-dependent mechanism.

dc.format.extent

4126327 bytes

dc.format.mimetype

application/pdf

dc.identifier.uri

https://hdl.handle.net/10161/1164

dc.language.iso

en_US

dc.subject

Biology, Genetics

dc.subject

Biology, Molecular

dc.subject

Biology, Microbiology

dc.subject

C

dc.subject

elegans

dc.subject

cep

dc.subject

nucleolus

dc.subject

p

dc.subject

Salmonella

dc.title

Inhibition of Nucleolar Proteins in Caenorhabditis Elegans Confers Enhanced Resistance to Salmonella Enterica through a P53/cep-1-Dependent Mechanism

dc.type

Dissertation

duke.embargo.months

24

Files

Original bundle

Now showing 1 - 1 of 1
Loading...
Thumbnail Image
Name:
D_Fuhrman_Laura_a_200904.pdf
Size:
3.94 MB
Format:
Adobe Portable Document Format

Collections