The RADI gene of Saccharomyces cerevisiae is involved in excision repair of damaged DNA. The nucleotide sequence of the RADI gene presented here shows an open reading frame of 3,300 nucleotides. Two ATG codons occur in the open reading frame at positions + 1 and +334, respectively. Since a deletion of about 2.7 kilobases of DNA from the 5' region of the RADI gene, which also deletes the +1 ATG and 11 additional codons in the RADi open reading frame, partially complements UV sensitivity of a rad1A mutant, we examined the role of the + 1 ATG and +334 ATG codons in translation initiation of RAD1 protein. Mutation of the + 1 ATG codon to ATC affected the complementation ability of the RADI gene, whereas mutation of the +334 ATG codon to ATC showed no discernible effect on RADI function. These results indicate that translation of RAD1 protein is initiated from the +1 ATG codon. Productive in-frame RADI-lacZ fusions showed that the RADI open reading frame is expressed in yeasts. The RAD1-encoded protein contains 1,100 amino acids with a molecular weight of 126,360. The RAD1 gene of Saccharomyces cerevisiae is one of 10 genes, RADI, RAD2, RAD3, RAD4, RAD7, RADIO, RAD14, RAD16, RAD23, and MMS19, involved in excision repair of DNA damaged with UV light, bulky adducts, or cross-linking agents (17-19, 30, 39). Of these, the RADI, RAD2, RAD3, RAD4, RADIO, and MMSJ9 genes seem to be required for incision of damaged DNA since their mutants are highly defective in this process, whereas the remaining four genes affect incision only partially (17,18,30,39). To analyze the individual and the combined role of proteins encoded by these incision genes, we have cloned and characterized several of them (10-12, 16, 20, 24, 25, 27, 28). Some of these genes have also been cloned and studied by others (21-23, 38, 40). The cloned genes are being used to overproduce their proteins to facilitate their purification, characterization, and assembly of the incision complex in vitro. Such an approach has allowed successful in vitro assembly and characterization of the UVRABC incision complex in Escherichia coli (34).As an important step toward achieving the above goals, we have examined the primary structure of the proteins encoded by the RAD2, RAD3, RAD7, and RADIO genes, as deduced from their nucleotide sequence (16,24,27,28). In the RAD3-encoded protein sequence we identified a segment which is homologous with a consensus sequence present in a large number of proteins which bind and hydrolyze ATP, suggesting that RAD3 protein also possesses a similar activity (27). The RADIO protein sequence indicates the presence of putative DNA-binding regions (28). Interestingly, significant amino acid sequence homology has been observed between the yeast RAD10 protein and the protein encoded by the human excision gene ERCC-J, suggesting that repair proteins in eucaryotes are conserved (37).In this paper, we present the nucleotide and the deduced amino acid sequences of RADI. The RADI open reading frame encodes a protein of 1,100 amino acids with a molec...
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
customersupport@researchsolutions.com
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.