Premium
Effects of ubiquitylation on the temporal progression of Nucleotide Excision Repair
Author(s) -
Lalimar Danielle,
Fischhaber Paula
Publication year - 2009
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.23.1_supplement.836.1
Subject(s) - nucleotide excision repair , yeast , mutant , ubiquitin , mutagenesis , nucleotide , plasmid , biology , saccharomyces cerevisiae , strain (injury) , microbiology and biotechnology , gene , genetics , dna repair , anatomy
This study is examining whether ubiquitylation of Rad4 influences the temporal progression of Nucleotide Excision Repair (NER). To investigate this, a Rad7SOCS mutant has been constructed in which amino acids L168 and C172 have been mutated to alanines thus rendering Rad7 deficient in its ubiquitylation capability. In this ubiquitylation deficient background it will be possible to determine if there is a change in the timed recruitment of NER and therefore if ubiquitylation regulates this timing. The Rad7SOCS mutant was prepared by site‐directed mutagenesis. The Rad7SOCS plasmid has been transformed into yeast and screening and sequencing efforts are currently underway to select for the Rad7SOCS yeast strain. After transforming into yeast, the Rad7SOCS strain will be combined genetically with strains containing the fluorescently labeled Rad10‐YFP and Rad14‐CFP genes. These strains, as well as the isogenic Rad7 wild type strain, will be UV‐irradiated and temporal changes in the appearance or disappearance of Rad10 and Rad14 will be observed.