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Phenotypic Characterization of LRS1‐2 Knockout Mutants in Arabidopsis thaliana
Author(s) -
Lozzi Brittney,
Keuls Rachel,
Erickson Patti,
Erickson F. Les
Publication year - 2016
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.30.1_supplement.1069.9
Subject(s) - mutant , biology , arabidopsis , arabidopsis thaliana , wild type , ubiquitin , ubiquitin ligase , genetics , saccharomyces cerevisiae , gene , microbiology and biotechnology , biochemistry
Ubiquitination is an important step in post‐translational protein modification and is tightly regulated within the cell. The Arabidopsis thaliana lateral root stimulator 1 ( LRS1 ) gene encodes a Damaged DNA Binding WD40‐repeat (DWD) protein that may play a role in ubiquination regulation. The DWD motif suggests an interaction with E3 ubiquitin ligases, but recent findings indicate that LRS1 may regulate deubiquitinases (DUBs), which reverse ubiquitination via cleavage of the bond between ubiquitin and its target protein. LRS1 is highly conserved among eukaryotic species, including humans, worms, and yeast, where it is involved in DNA repair, glutamate receptor abundance, and biosynthesis of mitochondrial subunits necessary for respiration. To determine its biological function in plants, T‐DNA insertion mutants of LRS1 have been identified. Initial observations of the lrs1‐1 allele revealed reduced lateral root growth with otherwise normal development. A second allele, lrs1‐2 , has been identified and genotyped using PCR. Homozygous lrs1‐ 2 mutants and their sibling, wild‐type segregants have been isolated after multiple rounds of back‐crossing mutants to the parental wild type Columbia (Col‐0). Both mutant lines are being analyzed for altered developmental and biological phenotypes, including germination rates, sensitivity to auxin and its transport, and reproductive capacity. lrs1 mutants are also being grown on media containing phenanthroline, isopropyl ( N ‐3‐chloro‐phenyl)‐carbamate, and nordihydroguaiaretic acid because Saccharomyces cerevisiae mutants lacking the LRS1 homolog, DUB‐associated factor 1 ( DUF1 ), are sensitive to these compounds. By exposing the mutant plant lines to different chemical and environmental stress conditions, a phenotype may be revealed that provides a better understanding of the cellular activities of LRS1 and its function in plants . Support or Funding Information This project is supported by an NSF Major Research Instrumentation grant and Salisbury University's Henson Undergraduate Research Fund.