Investigating the role of germline RNA helicase-1 in Caenorhabditis elegans

[ACCESS RESTRICTED TO THE UNIVERSITY OF MISSOURI AT AUTHOR'S REQUEST.] The germline specific RNA helicase GLH-1, a constitutive P-granule component, and DCR-1, the riboendonuclease responsible for processing small RNAs in the miRNA pathway, have been shown to be interdependent in the C. elegans germline. The work described here is based on the hypothesis that if GLH-1 functions with DCR-1 in the miRNA pathway in the C. elegans germline, then when GLH-1 is missing or reduced, there will be a difference in miRNA processing and/or abundance in glh-1 mutants as compared to wild type worms. We utilized deep sequencing on an Illumina platform to analyze differences in miRNA abundance between glh-1(gk100) mutant and wild type worms. In addition, to ascertain whether any miRNAs are germline specific, we also analyzed differences in miRNA expression between glp-4(bn2) worms cultured at both 15oC and 26oC. Results of deep sequencing and qRT-PCR indicated that in general miRNA abundance is moderately decreased in glh-1(gk100) mutants, and this reduction is significant in mutants cultured at 26oC. Our results also indicated that 11 of the 153 miRNAs identified to date in C. elegans are germline-enhanced, including miR-38, miR-40, miR-228, and miR-244, and may be germline-specific.

Name thesis:degree_name

Ph. D.

Level thesis:degree_level

Doctoral

Discipline thesis:degree_discipline

Molecular microbiology and immunology (MU)

Grantor dc:publisher

University of Missouri--Columbia

Year dc:date.issued

2011

Author dc:creator

• McEwen, Tamara J., 1965-

Advisor dc:contributor.advisor

• Bennett, Karen L.

• germline development
• miRNA pathway
• C. elegans

Statement dc:rights

• Access to files is limited to the campuses of the University of Missouri with SSO login.

Language dc:language.iso

eng, English

DOI

https://doi.org/10.32469/10355/14294

OAI identifier oai:identifier

oai:mospace.umsystem.edu:10355/14294