Helicases perform vital roles in the cell by unwinding D N A to make it accessible for the
essential processes of replication, transcription and repair. In Caenorhabditis elegans, the DOG-
1 helicase-like protein is required for polyG/polyC-tract (G/C-tract) maintenance, as dog-l
animals have a mutator phenotype characterized by deletions that initiate in G/C-tracts. DOG-1
may unwind secondary structures that form in polyguanine D N A during lagging strand
replication. In order to more completely understand the role of dog-1, genetic interactors were
identified, dog-1 functionally interacts with the him-6/BLM helicase. Absence of
recombinational repair-implicated proteins in the dog-1 background, including HIM-6/BLM,
RAD-51, BRD-1/BARD1 and HIM-9/XPF, as well as the trans-lesion synthesis polymerases
polKMD po/7 increased the frequency of animals with G/C-tract deletions, indicating that these
pathways are important mechanisms for repair at G/C-tracts in the absence of DOG-1. These
data support the hypothesis that persisting D N A secondary structures can cause replication fork
stalling, which can be resolved by deletion-free or deletion-prone mechanisms.
DOG-1 has highest sequence identity to human BR1P1/FANCJ, which is mutated in
patients from the Fanconi Anemia (FA) subgroup J. D N A damage sensitivity experiments
indicated that, like chicken F A N C J cells, dog-1 mutants were not significantly sensitive to DNA
damage from X-ray or UV-irradiation, but were extremely hypersensitive to the D N A interstrand
cross-linking agent UVA-activated trimethylpsoralen. Thus, DOG-1 appears to have a
conserved role in cross-link repair and is the C. elegans F A N C J homolog. Characterization of
the dog-1/FANCJ-relatsd helicase, Jrh-1, revealed that mutants for this putative helicase are
moderately sensitive to cross-linking agents, dog-1 jrh-1 double mutants displayed a synthetic
lethal phenotype characterized by excessive recombination intermediates and mitotic catastrophe
in the germline. However, absence of JRH-1 did not have any effect on G/C-tract deletions,
indicating that JRH-1 does not have a redundant function with DOG-1 at G/C-tracts. Absence of
JRH-1 reduced the fitness of eTl and nTl translocation hétérozygotes, but not translocation
homozygotes, jrh-1 was synthetically lethal with him-6/BLM and with the endonuclease mus-81,
suggesting a possible role for JRH-1 in regulating the balance between different types of repair. / Medicine, Faculty of / Medical Genetics, Department of / Graduate
| Identifer | oai:union.ndltd.org:UBC/oai:circle.library.ubc.ca:2429/2832 |
| Date | 05 1900 |
| Creators | Youds, Jillian L. |
| Publisher | University of British Columbia |
| Source Sets | University of British Columbia |
| Language | English |
| Detected Language | English |
| Type | Text, Thesis/Dissertation |
| Format | 11279819 bytes, application/pdf |
| Rights | Attribution-NonCommercial-NoDerivatives 4.0 International, http://creativecommons.org/licenses/by-nc-nd/4.0/ |
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