ISOLATING THE TARGETS OF SIX TRANSCRIPTION FACTOR IN EPHYDATIA MUELLERI AND IDENTIFYING THE ROLE OF THE SUPEROXIDE DISMUTASE 6 IN HOST IMMUNE RESPONSE TO TRICHOMONAS VAGINALIS

Author

Gurbir Kaur Gudial, University of the PacificFollow

Date of Award

2017

Document Type

Thesis

Degree Name

Master of Science (M.S.)

Department

Biological Sciences

First Advisor

Lisa Wrischnik

First Committee Member

Ajna Rivera

Second Committee Member

Kirkwood Land

Abstract

Sponges are the descendants of the oldest members of the metazoan phylogenetic lineage and their genome contains animal specific genes but lack true tissues, organ systems, and neurons. Thus, the sponge model system can be used to elucidate origin of developmental processes. The PSED (RDGN) network (Pax/Six/Eya/Dac) is important in development of eyes, muscles, and other structures in Bilaterians. Similarly, sponges contain a precursor Pax-Six gene network. The Ephydatia muelleri (Em) PaxB protein binds to a

Pax2/5/8 consensus sequence site and two cis-regulatory elements upstream and one intron sequence of EmSix1/2 (Rivera et al., 2013).

This study aimed to determine if transcription factor EmSix1/2 binds upstream of EmPaxB using gel shift mobility assays, identify other downstream targets of EmSix1/2 using DNA immunoprecipitation, and to identify the recognition sequence of Six in sponges through sequencing. In conclusion, purified EmSix binds to DNA specific fragments (1 and 3), which may contain enhancer sequences located in the PaxB promoter region.Possible consensus recognition sequence of Six in sponges were also identified.

The host immune response has various mechanisms to protect the organism from infections and invasions of microorganisms and cell damaging chemicals. One such mechanism is the elimination of reactive oxygen species aided by superoxide dismutase (SOD). A study showed that anti-neutrophil chemotactic factor antibodies recognize Tritrichomonas foetus SOD (Granger et al., 1997). During first casualties, SOD is released and triggers a host immune response. The parasites could use SOD to counter oxidative attacks by the leukocytes and damaged cells to protect surrounding parasites. We used the parasite Trichomonas vaginalis that causes trichomoniasis to determine if SOD acts a neutrophil (or other leukocyte) chemotactic factor in this parasite and characterize the expression and secretions of epithelial cells when treated with SOD6. HeLa cells treated with SOD6 showed an increase in the expression of IL-8 chemokine relative to TrxR treated cells. Scratch test assays showed that SOD may act as a macrophage chemoattractant as compared to TrxR but further tests are needed to confirm this.

Recommended Citation

Gudial, Gurbir Kaur. (2017). ISOLATING THE TARGETS OF SIX TRANSCRIPTION FACTOR IN EPHYDATIA MUELLERI AND IDENTIFYING THE ROLE OF THE SUPEROXIDE DISMUTASE 6 IN HOST IMMUNE RESPONSE TO TRICHOMONAS VAGINALIS. University of the Pacific, Thesis. https://scholarlycommons.pacific.edu/uop_etds/2972