Date of Award
Master of Science
Dr. Susanne Kohalmi
Dr. Rima Menassa
The enzyme arogenate dehydratase (ADT) operates in phenylalanine biosynthesis. To determine the substrate specificity and subcellular localization of all six Arabidopsis ADTs, pha2 yeast complementation assays and confocal microscopy analyses were performed. The pha2 yeast complementation assay was used to determine the ADTs that can accept prephenate as a substrate in vivo. Results show that ADT1, ADT2 and ADT4 were able to complement the pha2 phenotype, but PDT function is temperature dependent. Results from confocal microscopy analyses show that ADT-CFP fusion proteins including transit peptide sequences are targeted to chloroplasts (ADT1-ADT5), the nucleus (ADT5) or remain cytosolic (ADT6). Immunoblot analyses show that all ADT transit peptides are cleaved in planta. Together these results show differences in the substrate specificity and subcellular localization ofArabidopsis ADT isoforms.
Bross, Crystal Dawn, "CHARACTERIZING SUBSTRATE SPECIFITY AND SUBCELLULAR LOCALIZATION OF AROGENATE DEHYDRATASES FROM ARABIDOPSIS THALIANA" (2011). Digitized Theses. 3380.