PgmNr Y3005: The Yeast Genome Project: Exploring APD1.

Authors:
C. A. Douds; J. Keeney


Institutes
Juniata College, Huntingdon, PA.


Keyword: Cell Cycle/Growth Control/Metabolism

Abstract:

Yeast is considered to be “man’s oldest industrial microorganism” and has been used by humans for over 5,000 years. Today, its use has expanded and Saccharomyces cerevisiae has become an ideal eukaryotic model due to its quick growth and easily manipulated genome. After its genome was sequenced in 1996, scientists came together to create a full encyclopedia of systemized gene functions characterized according to the Gene Ontology Consortium (GO). However, nearly twenty years later, the functions of more than 10% of these genes still remain unknown. The ORFan gene project aims to aid the characterization of the S. cerevisiae genome by exploring the function of specific open reading frames (ORFs). In order to classify these unknown ORFs, a putative pipeline has been constructed beginning with the identification of an ORFan of interest through the Saccharomyces Genome Database (SGD). This project focuses on YBR151W or APD1. The selected ORF is fluorescently tagged at the C and N terminals by PCR and visualized with fluorescent microscopy to verify cellular localization. Then a deletion strain is constructed and SGD reported phenotypes are verified. Verified phenotypes for Δapd1 cells include increased H2O2 sensitivity and abnormal actin patch localization. Next, a comparison between the wild type and target gene knockout strains is analyzed through RNAseq. However, in this study RNAseq data has yet to be obtained. Instead, microarray analysis using SGD’s SPELL was conducted to explore hypotheses. Further exploration into the putative function of APD1 has led to studies involving actin patch localization, as well as the protein’s potential role in oxidative stress mechanisms. We have observed the effects of actin patch localization under various forms of oxidative stress such as HU and H2O2 between wildtype and Δapd1 strains.



Yeast Database Genetic Index
1. gene symbol: APD1; systematic name: YBR151W