Use of in vivo and in vitro assays for the characterization of mammalian excision repair and isolation of repair proteins.
Elucidation of the molecular mechanism of mammalian nucleotide excision repair requires the availability of purified proteins, DNA substrates with defined lesions and suitable repair assays. Repair assays introduced in recent years vary from testing individual steps and successions of steps in vitro to systems that closely reflect the entire process in vivo. In the first part of this review, an in vivo microinjection system is discussed. The second part of the article reviews an in vitro system for study of repair synthesis promoted by cell extracts. Both systems can be utilized as assays during the purification of protein factors that complement repair-defective xeroderma pigmentosum cells. The effect of purified repair proteins from other organisms on mammalian repair is also considered.
Department of Cell Biology and Genetics, Erasmus University, Rotterdam, The Netherlands.
J H
JH
Hoeijmakers
A P
AP
Eker
R D
RD
Wood
P
P
Robins
D000818:Animals; D001681:Biological Assay / Q000379:methods; D002457:Cell Extracts / Q000494:pharmacology; D004247:DNA / Q000096:biosynthesis*; D004260:DNA Repair / Q000187:drug effects / Q000528:radiation effects; D004305:Dose-Response Relationship, Drug; D004307:Dose-Response Relationship, Radiation; D006801:Humans; D066298:In Vitro Techniques; D008730:Methoxsalen / Q000494:pharmacology; D008845:Microinjections; D010957:Plasmids; D010984:Platinum / Q000494:pharmacology; D014466:Ultraviolet Rays / Q000009:adverse effects
D002457:Cell Extracts; D010984:Platinum; D004247:DNA; D008730:Methoxsalen