Studying the Role of Prenylated Rab Acceptor 1 Domain Family, Member 2 (PRAF2) in Drosophila melanogaster by Knockdown and Gain-of-Function Analysis
Studying the Role of Prenylated Rab Acceptor 1 Domain Family, Member 2 (PRAF2) in Drosophila melanogaster by Knockdown and Gain-of-Function Analysis
Date
2016-05
Authors
Xu, Zhaotong
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de Couet, Heinz Gert
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Molecular Cell Biology
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University of Hawaii at Manoa
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Abstract
Prenylated Rab acceptor family (PRAF) proteins are highly conserved among multicellular organisms and several paralogous genes of PRAF exist. PRAF proteins are expressed in a variety of tissues and are associated with cellular transport and endo/exocytic vesicle trafficking via Rab protein interactions. Currently, the function of a new PRAF protein, PRA1 domain family, member 2 (PRAF2) is still not understood. However, PRAF2 is upregulated in cancerous cells of the breast, colon, lung, and ovaries and serves as a candidate prognostic marker for neuroblastoma. This study focuses on studying the role of PRAF2 in Drosophila melanogaster by knockdown and gain-of-function analysis. In the first part of the experiment, a knockdown of PRAF2 in Drosophila ommatidia was achieved using RNAi methodologies. Results for the PRAF2 knockdown showed that PRAF2 expression at 25°C was sufficient in causing structural abnormalities in Drosophila ommatidia. When flies were grown at 28°C, results were inconclusive because the specific and non-specific effects of the GMR-GAL4 driver and PRAF2-RNAi could not be distinguished. In the second part of the experiment, PRAF2 was overexpressed in Drosophila. Fly eye specimens were then obtained for further examination using transmission electron microscopy (TEM) and scanning electron microscopy (SEM). Results showed that PRAF2 overexpression correlated with increased spacing between photoreceptor cells in Drosophila ommatidia, which also affected ommatidia shape. Further research must be conducted on PRAF2 to better understand its mechanisms in vesicular trafficking, which might provide insight on how abnormalities in PRAF2 function can contribute towards the development of cancerous cells.
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Prenylated Rab acceptor family (PRAF) proteins,
endocytic/exocytic vesicle trafficking,
cellular transport,
Rab proteins,
PRA1 domain family,
member 2 (PRAF2)
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34 pages
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