In Vitro Correction of Cells from Patients with Mutagen Hypersensitivity
An important strategy in the study of mutant cells, such as those from patients with an inherited hypersensitivity to UV or X-rays, is based on the principle of correction. In this contribution various approaches of in vitro correction are illustrated with complementation analysis of ataxia telangiectasia (AT) and with the introduction of wild-type protein extracts or yeast photoreactivating enzyme into xeroderma pigmentosum (XP) cells.
The combined results from genetic complementation studies in X-ray sensitive patients using the radioresistant DNA synthesis as a marker indicate the existence of six different complementation groups. Four of these (AB, C, D and E) are represented by AT patients and two (V1 and V2) harbour patients with the “Nijmegen Breakage syndrome”. A seventh separate category is formed by a number of AT patients with a normal inhibition of DNA synthesis by X-rays.
In XP cells a transient correction of the defect was accomplished by micro-injection of cell-free extracts from wild-type sources. Some properties of the factor correcting XP cells from complementation group A are described.
Micro-injection was also used to introduce the yeast photo-reactivating enzyme (PRE) into cells. After light-induced removal of pyrimidine dimers in injected cells the levels of UV-induced unscheduled DNA synthesis (UV-UDS) decreased only slowly. Kinetic studies suggest, that the average processing time of dimer excision is less than 1 hour in normal cells and more than 3 hours in some XP complementation groups.
KeywordsAtaxia Telangiectasia Average Processing Time Nijmegen Breakage Syndrome NIJMEGEN Breakage Syndrome Patient Mutagen Hypersensitivity
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