Molecular Biology Basis for the Response of Poly (ADP-RIB) Polymerase and NAD Metabolism to DNA Damage Caused by Mustard Alkylating Agents
Abstract
During the midterm of this contract we have performed a variety of experiments to provide a strategy to modulate the nuclear enzyme poly (ADP- ribose) polymerase, in cultured keratinocytes. This enzyme modifies a variety of nuclear proteins utilizing NAD. DNA is required for the catalytic activity of the enzyme and the activity is dependent upon the presence of strand breaks in this DNA. It has been hypothesized that human skin exposed to mustards may develop blisters due to a generalized lowering of NAD in exposed skin cells. Furthermore, one mechanism may be the production of DNA strand breaks in skin cells by mustards which activate NAD depletion in these cells via the poly(ADP- ribose) polymerase reaction as described above. During the midterm we have cloned various regions of the cDNA for poly(ADP-ribose) polymerase into a series of retroviral expression vectors. We have shown that when keratinocytes are transfected with such expression vectors, increased amounts of poly(ADP-ribose) polymerase mRNA and protein are expressed. The projects are currently exploring the use of a new retroviral system (N2) which offers several advantages over the retroviral expression system system used earlier. Retroviral expression vectors should allow us to modulate the activity of this nuclear enzyme in a variety of skin cells in vitro. and in vivo. Keywords: RA V; Human tissue; Skin; Lab animals; Cell cultures; Athymic mice; Sulfur mustard; Poly(ADP-Ribese).
Document Details
- Document Type
- Technical Report
- Publication Date
- Aug 01, 1988
- Accession Number
- ADA218313
Entities
People
- Mark E. Smulson
Organizations
- Georgetown University