The Yeast Genome Project: Characterizing APD1

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 was fluorescently tagged at the C and N terminals to verify cell localization. Next, a comparison between the wild type and target gene knockout strains will be analyzed through RNAseq. This comparative analysis uses programs such as FastQC, FastX, RSEM, Bowtie2 and R, and will provide information about the impact of gene deletion on global gene expression levels. 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 H 2 O 2 between wildtype and Δapd1 strains. Support or Funding Information Howard Hughes Medical Institute Undergraduate Grant Program, Funded by HHMI GCAT SEEK SNF grant #1248096, NIH Grant R15‐GM0587 to Jill B. Keeney