Date of Award
Winter 1-21-2026
Degree Type
Thesis
Degree Name
Master of Science (MS)
Department
Biology
First Advisor
Stefan Kanzok
Abstract
Investigating the role of PhLP3 in the fecundity of the malaria vector Anopheles stephensi Phosducin-like proteins (PhLPs) are small thioredoxin domain-containing proteins that are highly conserved in eukaryotes. PhLPs are subdivided into three subclasses, each playing different cellular roles, from modulating G-protein signaling (PhLP1) to regulating the actin and tubulin cytoskeleton (PhLPs2&3). In a collaborative project, we recently discovered that Drosophila flies lacking PhLP3 are sterile. We hypothesize that the same may be true for the malaria-transmitting mosquito, Anopheles stephensi. Here, I present the identification and biochemical characterization of A. stephensi PhLP3 (AsPhLP3). Sequence and structural analysis of AsPhLP3 shows the characteristic organization of the PhLP family with an N-terminal helix domain, a central thioredoxin domain, and a short C-terminal tail domain. I confirmed AsPhLP3 gene expression in the reproductive tissues of Anopheles via RT-qPCR. Then, I cloned, expressed, and purified recombinant AsPhLP3. Initial insulin reduction assays confirm redox activity, as previously demonstrated for Drosophila PhLP3. For the first time, I show tissue-specific protein localization of a PhLP using anti-AsPhLP3-specific antibodies. I demonstrate that PhLP3 is expressed in the reproductive organs of the mosquito, where it appears to co-localize with microtubules. With the goal of generating an A. stephensi line lacking PhLP3, I acquired and generated all of the components for the mosquito-specific CRISPR/Cas9 system called ReMOT. I injected 160 mosquitoes, but I was unsuccessful in detecting editing events in vivo. Anopheles mosquitoes are among the most prominent global disease vectors, transmitting infectious diseases such as Malaria and Dengue Fever. New insights into the function of AsPhLP3 and its potential involvement in mosquito fecundity may open new avenues to combat this critical disease vector.
Recommended Citation
Webster, Samantha Rose, "Intital Characterization of Phosducin-Like Protein 3 in the Malaria Vector Anopheles Stephensi" (2026). Dissertations. 4279.
https://ecommons.luc.edu/luc_diss/4279
