Date Completed: 2/5/08
Faculty Name: Laura Timares
UAB Department: Dermatology, Division of Human Gene Therapy
UAB School: School of Medicine
Campus Address: BMR2-542
Campus Zip: 2172
Email Address: firstname.lastname@example.org
Research Program Area: Tumor Biology
Project Title: The role of Bid in promoting cell repair in response to UV-damage
Project Status: Already up-and-running
Proposed Start Date: April 14
Proposed End Date: July 25
Number of Weeks of Internship: 15
Number of Interns: 1
Other faculty, staff, or graduate students who may help supervise intern: Hee Kyung Kim
Expected Number of Work Hours Per Week: 37.5
Expected Work Schedule for Intern: Flexible, intern can largely set his or her own schedule
Category of Research: Laboratory Research
Cancer Research: Skin
Project Description: DNA damage by the sun activates a complex cellular reponse that must determine the cell fate. The molecular sensors that regulate this fate can orchestrate events that promote cellular repair or cellular suicide called apoptosis. The barrier function of the skin is required for survival of an organism, and so the sun exposed epidermal cells, called keratinocytes, are highly evolved in making cellular decisions that promote barrier function at the risk of incorporating low levels of mutations that may later develop cancer. A recognized regulator of this decision making process is the tumor supressor p53 molecule. We propose that the pro-apoptotic molecule, Bid, may also act as an important cellular sensor and promote either cell survival or death. Recent evidence has revealed that a prosurvival isoform of Bid is important for cell cycle check-point function in response to gamma irradiation, but its role in UV responses is not understood. We have observed that UV exposure induces a nuclear translocation of Bid, and that Bid is phosphorylated at a site that is recognized by the DNA damage sensor, ATR, generating the pro-survival isoform (pBid). A CaRES student may work on one of the following aims:
1) To determine if ATR is indeed responsible for generating pBid, by examining the impact of drug and genetic inhibitors. Development of a lentiviral construct encoding mouse and/or human ATR inhibitory short hairpin RNA will be generated for that purpose.
2) To determine what the consequences are on cell cycle in wild type and Bid deficient keratinocytes derived from knock-out mice and on deficient cells reconstitute with WT and mutant of Bid genes. Lentivirus DNA constructs encoding these genes are in hand.
3) To determine the role of Bid in UV damaged human skin cancer cells and the requirement for p53. Parental p53 deficient and transfected p53 positive lines will be examined for pBid generation in response to UV light. Lentivirus encoding Bid specific inhibitory shRNA will be generated and used to infect these human tumor lines. The absence of Bid expression will be confirmed by RT-PCR and immunostaining. Then UV-induced cell-cycle checkpoint function will be examined.
Intern's Anticipated Duties:
Duty 1: Perform experiments involving isolation and culture of mouse skin cells. Immunoflourescence staining, BrdU-based cell cycle assays, flow cytometry
Duty 2: Molecular biology in developing lentiviral constructs, transfection procedures and selection of cell clones
Duty 3: Keeping a complete lab notebook and write-up of the project
Preceptor will provide intern with access to the following: office or desk space; computer and printer; laboratory work bench space; equipment needed to complete project; supplies needed to complete project
Likelihood of authoring publications: Very likely
Background, education, experience, or expertise preferred: Animal Research; Cell Biology; Genetics; Infetious Agents and Cancer; Basic Knowledge of Lab Skills; Advanced Knowledge of Lab Skills; Literature Review Skills; Molecular Biology; Scientific Writing Skills; Basic Knowledge of Statistics and Data Management
This faculty member has been a CaRES Preceptor for three or more summers.
Intern 1: open