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Essential functions of IFA-2 domains in Caenorhabditis elegans fibrous organelles

Williams, Kyle C.
http://rave.ohiolink.edu/etdc/view?acc_num=toledo1333748571

Abstract Details

2012, Master of Science, University of Toledo, Biology (Cell-Molecular Biology).
The C. elegans fibrous organelle (FO) consists of apical and basal hemidesmosomes linked by cytoplasmic intermediate filaments (IFs). FOs are most prominent in the epidermis where they overlie body wall muscle and serve to transmit force from the muscle to the cuticle. IFA-2, one of four epidermally expressed IFs whose loss results in epidermal fragility and failure of muscle-cuticle force transmission, localizes to FOs. All IFs contain three domains: a central rod domain and globular head and tail domains. The rod is essential for assembly of IFs into mature filaments while less is known about the functions of the head and tail domains. Roles of the head and tail domains of IFA-2 were examined by expressing GFP-tagged IFA-2 variants in transgenic animals and examining their phenotype and localization in wild-type and null backgrounds. An IFA-2 variant containing only the head domain (IFA-2H) was able to localize to FOs on its own. High levels of larval-stage IFA-2H incorporation into FOs correlate with a dominant-negative muscle detachment phenotype, while low-level incorporation results in healthy animals. This suggests the head domain interacts with FO components essential to tissue integrity. To determine possible interactions between the head domain and FO proteins, the yeast two-hybrid system was used. An initial screen detected that the head and tail domains of IFA-2 are able to interact with each other. To obtain a more complete list of IFA-2H interactors, a C. elegans cDNA library was screened against IFA-2H. After PCR and sequencing of the 239 clones generated from the library screen, 45 unique proteins were found to interact with IFA-2H. Several criteria including functional roles, tissue expression data, and frequency of library inserts has been used to analyze the data and assign relevance to the interactions. One of the proteins found in the library screen, K04G7.1, was investigated further and found to localize throughout the pharynx where it may interact with IFs. This work should help to elucidate the role of the different domains of IFA-2 in FO function and reveal the underlying defects leading to tissue fragility.
John Plenefisch, Ph.D (Committee Chair)
Bruce Bamber, Ph.D (Committee Member)
Scott Leisner, Ph.D (Committee Member)
Robert Steven, Ph.D (Committee Member)
84 p.
Developmental Biology
C. elegans; Intermediate Filaments

Recommended Citations

Citations

  • Williams, K. C. (2012). Essential functions of IFA-2 domains in Caenorhabditis elegans fibrous organelles [Master's thesis, University of Toledo]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=toledo1333748571

    APA Style (7th edition)

  • Williams, Kyle. Essential functions of IFA-2 domains in Caenorhabditis elegans fibrous organelles. 2012. University of Toledo, Master's thesis. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=toledo1333748571.

    MLA Style (8th edition)

  • Williams, Kyle. "Essential functions of IFA-2 domains in Caenorhabditis elegans fibrous organelles." Master's thesis, University of Toledo, 2012. http://rave.ohiolink.edu/etdc/view?acc_num=toledo1333748571

    Chicago Manual of Style (17th edition)

toledo1333748571
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© 2012, all rights reserved.
This open access ETD is published by University of Toledo and OhioLINK.