Honors Theses
Date of Award
Spring 5-8-2025
Document Type
Undergraduate Thesis
Department
Biology
First Advisor
Bradley Jones
Second Advisor
Sarah Liljegren
Third Advisor
Mika Jekabson
Relational Format
Dissertation/Thesis
Abstract
In Drosophila melanogaster, the mechanism influencing the differentiation of glia and neurons is directed by a key transcription factor known as glial cells missing (gcm). gcm is a crucial regulator of cell fate decision, functioning as a binary switch to direct glial fate in gcm- positive cells and neuronal fate in gcm-negative cells (Jones, et.al, 1995; Hosoya, et.al, 1995; and Vincent, et.al, 1996). In addition to directing gliogenesis, gcm and its homolog gcm2 facilitate the determination of macrophages and tendon cells, highlighting that gcm acts with supplementary factors to induce various signaling pathways in different contexts (Jones, 2005; Johnson, et al., 2012). Glia development is initiated when Gcm activates downstream factors like Repo in the terminal embryonic phase of Drosophila. Repo cooperates alongside transcription factors such as Pointed p1 for migration and differentiation of glial cells and Tramtrack p69 for neuronal development inhibition (Yuasa et al., 2003; Lee and Jones, 2005; Johnson, et al., 2012). Repo expression is regulated through a critical 4.2 kb DNA area upstream of the repo gene called the repo cis-regulatory DNA region (Lee & Jones, 2005). Preliminary studies have identified a 98 bp fragment within the repo cis-regulatory region containing a single Gcm binding site (GBS) through which cofactors interact to modulate glial expression. (Lee & Jones, 2005; Johnson, et al., 2012). One such coregulator is Gro which belongs to a family of co-repressors and is recruited by various transcription factors, such as Gcm, to direct cell development (Kaul, 2014). It may be possible that Gro interacts with Gcm to regulate glial cell determination, yet the exact nature of their interaction remains unclear. To identify whether Gro directly or indirectly binds with Gcm through GBS, this study aims to co-express Gcm and Gro in the S2 cell line. Due to the lack of a suitable antibody for Gcm detection, a Myc tag was added to Gcm creating two different versions of the target protein: Myc-Gcm N-term and Myc-Gcm C-term. Subsequent staining of Myc and Gro in the western blot analysis detected the presence of the proteins before co-immunoprecipitation. This protocol is now available in the Jones lab to investigate potential protein candidates inducing gliogenesis with co-immunoprecipitation confirming the interaction.
Recommended Citation
Kosalram, Nivedha, "A Search for Transcriptional Cofactors of Gcm/Repo involved in Determining Glia Cell Fate in Drosophila melanogaster" (2025). Honors Theses. 3303.
https://egrove.olemiss.edu/hon_thesis/3303
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