Cloning, comparative mapping, and RNA expression of the mouse homologues of the Saccharomyces cerevisiae nucleotide excision repair gene RAD23

• Published in: Genomics (San Diego, Calif.) Calif.), 1996, Vol.31 (1), p.20-27
• Main Authors: VAN DER SPEK, P. J, VISSER, C. E, HANAOKA, F, SMIT, B, HAGEMEIJER, A, BOOTSMA, D, HOEIJMAKERS, J. H. J
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Animals; Biological and medical sciences; Chromosome Mapping; Cloning, Molecular; Conserved Sequence; DNA Repair - genetics; DNA Repair Enzymes; DNA, Complementary - genetics; DNA-Binding Proteins - genetics; Evolution, Molecular; Fundamental and applied biological sciences. Psychology; Fungal Proteins - genetics; Gene Expression - radiation effects; Genes, Fungal; Genes. Genome; Humans; In Situ Hybridization, Fluorescence; Male; Mice; Molecular and cellular biology; Molecular genetics; Molecular Sequence Data; Multigene Family; RNA, Messenger - genetics; RNA, Messenger - metabolism; Saccharomyces cerevisiae - genetics; Saccharomyces cerevisiae Proteins; Sequence Homology, Amino Acid; Species Specificity; Testis - metabolism; Ubiquitins - genetics; Ultraviolet Rays
• ISSN: 0888-7543; 1089-8646
• DOI: 10.1006/geno.1996.0004

The Saccharomyces cerevisiae RAD23 gene is involved in nucleotide excision repair (NER). Two human homologs of RAD23, HHR23A and HHR23B (HGMW-approved symbols RAD23A and RAD23B), were previously isolated. The HHR23B protein is complexed with the protein defective in the cancer-prone repair syndrome xeroderma pigmentosum, complementation group C, and is specifically involved in the global genome NER subpathway. The cloning of both mouse homologs (designated MHR23A and MHR23B) and detailed sequence comparison permitted the deduction of the following overall structure for all RAD23 homologs: an ubiquitin-like N-terminus followed by a strongly conserved 50-amino-acid domain that is repeated at the C-terminus. We also found this domain as a specific C-terminal extension of one of the ubiquitin-conjugating enzymes, providing a second link with the ubiquitin pathway. By means of in situ hybridization, MHR23A was assigned to mouse chromosome 8C3 and MHR23B to 4B3. Because of the close chromosomal proximity of human XPC and HHR23B, the mouse XPC chromosomal location was determined (6D). Physical disconnection of the genes in mouse argues against a functional significance of the colocalization of these genes in human. Northern blot analysis revealed constitutive expression of both MHR23 genes in all tissues examined. Elevated RNA expression of both MHR23 genes was observed in testis. Although the RAD23 equivalents are well conserved during evolution, the mammalian genes did not express the UV-inducible phenotype of their yeast counterpart. This may point to a fundamental difference between the UV responses of yeast and human. No stage-specific mRNA expression during the cell cycle was observed for the mammalian RAD23 homologs.