THE N-DEGRON PROTEIN-DEGRADATION STRATEGY FOR INVESTIGATING THE FUNCTION OF ESSENTIAL GENES - REQUIREMENT FOR REPLICATION PROTEIN-A AND PROLIFERATING CELL NUCLEAR ANTIGEN PROTEINS FOR NUCLEOTIDE EXCISION-REPAIR IN YEAST EXTRACTS

Nucleotide excision repair (NER) of DNA in the yeast Saccharomyces cer evisiae and in human cells has been shown to be a biochemically comple x process involving multiple gene products. In yeast, the involvement of the DNA replication accessory proteins, replication protein A (RPA1 ) and proliferating cell nuclear antigen (PCNA) in NER has not been de monstrated genetically. In this study we have generated temperature-de gradable rfa1 and pcna mutants and show that these mutants are defecti ve in NER in vitro under conditions that promote degradation of the RF A1 and PCNA gene products. We also demonstrate a physical interaction between RPA1 protein and subunits of the RNA polymerase II basal trans cription factor IIH (TFIIH). (C) 1998 Elsevier Science B.V. All rights reserved.