Chromosomal Localization and cDNA Cloning of the Genes (DDB1 and DDB2) for the p127 and p48 Subunits of a Human Damage-Specific DNA Binding Protein

doi:10.1006/geno.1995.1215

Genomics

Volume 29, Issue 1, 1 September 1995, Pages 62-69

https://doi.org/10.1006/geno.1995.1215 Get rights and content

Abstract

DDB is a damage-specific DNA binding protein whose binding activity is absent from a minority of cell strains from individuals with xeroderma pigmentosum Group E, a human hereditary disease characterized by defective nucleotide excision DNA repair and an increased incidence of skin cancer. The binding activity from Hela cells is associated with polypeptides of M_r 124,000 and 41,000 as determined by SDS-polyacrylamide gels. This report describes the isolation of full-length human cDNAs encoding each polypeptide of DDB. The predicted peptide molecular masses based on open reading frames are 127,000 and 48,000. When expressed in an in vitro rabbit reticulocyte system, the p48 subunit migrates with an M_r of 41 kDa on SDS-polyacrylamide gels, similarly to the peptide purified from Hela cells. There is no significant homology between the derived p48 peptide sequence and any proteins in current databases, and the derived peptide sequence of pl27 has homology only with the monkey DDB p127 (98% nucleotide identity and only one conserved amino acid substitution. Using a fluorescence in situ hybridization technique, the DDB p127 locus (DDB1) was assigned to the chromosomal location 11q12-q13, and the DDB p48 locus (DDB2) to 11p11-p12.

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Our results revealed that DDB1 expression is heterogeneous in ovarian cancer, suggesting its use as a potential biomarker for poor survival in ovarian cancer.
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UV-damaged DNA binding protein (UV-DDB) is a heterodimeric complex, composed of DDB1 and DDB2, and is involved in global genome nucleotide excision repair. Mutations in DDB2 are associated with xeroderma pigmentosum complementation group E. UV-DDB forms a ubiquitin E3 ligase complex with cullin-4A and RBX that helps to relax chromatin around UV-induced photoproducts through the ubiquitination of histone H2A. After providing a brief historical perspective on UV-DDB, we review our current knowledge of the structure and function of this intriguing repair protein. Finally, this article discusses emerging data suggesting that UV-DDB may have other non-canonical roles in base excision repair and the etiology of cancer.
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This specialized lesion recognition process, involving the interplay between XPC and UV-DDB, is intricately regulated by posttranslational protein modifications such as ubiquitination. The human DDB2 gene, located on chromosome 11 (11p11.2), consists of 10 exons encoding a 48 kDa protein of 427 amino acids [154]. Orthologs are present in many vertebrates, but are absent in other eukaryotes such as yeasts.
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The UV-damaged DNA-binding protein complex (UV-DDB) is involved in the damage recognition step of NER (Fig. 1). UV-DDB consists of two tightly associated subunits, DDB1 and DDB2.56–58 UV-DDB exhibits very strong and specific binding to UV-treated DNA and plays a role in GG-NER.59,60
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Regular ArticleChromosomal Localization and cDNA Cloning of the Genes (DDB1 and DDB2) for the p127 and p48 Subunits of a Human Damage-Specific DNA Binding Protein

Abstract

Regular Article
Chromosomal Localization and cDNA Cloning of the Genes (DDB1 and DDB2) for the p127 and p48 Subunits of a Human Damage-Specific DNA Binding Protein