Lack of transcription-coupled repair in mammalian ribosomal RNA genes.

We studied the induction and removal of UV-induced cyclobutane pyrimidine dimers (CPDs) in the ribosomal RNA genes (rDNA) in cultured hamster and human cells. In these genes, which are transcribed by RNA polymerase I, we found no evidence for transcription-coupled repair. The induction of CPDs was heterogeneous in rDNA due to nucleotide sequence: it was lower on the transcribed strand than on the nontranscribed strand and slightly lower in the coding region than in the nontranscribed spacer. Nevertheless, no dramatic difference in CPD induction was observed between rDNA and the dihydrofolate reductase (DHFR) gene. In Chinese hamster ovary cells, we observed no removal of CPDs from either rDNA strand within 24 h after UV irradiation. In these experiments, we did observe efficient repair of the transcribed, but not the nontranscribed, strand of the DHFR gene, in agreement with published results. In human cells, repair of rDNA was observed, but it showed no strand preference and was slower than that reported for the genome overall. No significant differences in repair were observed between restriction fragments from transcribed and nontranscribed regions or between growth-arrested and proliferating human cells, with presumably different levels of transcription of rDNA. We conclude that the modest level of rDNA repair is accomplished by a transcription-independent repair system and that repair is impeded by the nucleolar compartmentalization of rDNA. We discuss the possibility that recombination, rather than repair, maintains the normal sequence of rDNA in mammalian cells.

[1]  A. Sancar,et al.  Molecular mechanism of transcription-repair coupling. , 1993, Science.

[2]  P. Ghanouni,et al.  The RNA polymerase II elongation complex. Factor-dependent transcription elongation involves nascent RNA cleavage. , 1992, The Journal of biological chemistry.

[3]  P. Hanawalt,et al.  Inhibition of transcription and strand-specific DNA repair by alpha-amanitin in Chinese hamster ovary cells. , 1992, Mutation research.

[4]  H. Noller,et al.  Unusual resistance of peptidyl transferase to protein extraction procedures. , 1992, Science.

[5]  J. B. Walsh,et al.  Biased gene conversion, copy number, and apparent mutation rate differences within chloroplast and bacterial genomes. , 1992, Genetics.

[6]  A. Sancar,et al.  Escherichia coli mfd mutant deficient in "mutation frequency decline" lacks strand-specific repair: in vitro complementation with purified coupling factor. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[7]  P. Hanawalt,et al.  The genetic defect in the Chinese hamster ovary cell mutant UV61 permits moderate selective repair of cyclobutane pyrimidine dimers in an expressed gene. , 1991, Mutation research.

[8]  L. Mullenders,et al.  Xeroderma pigmentosum complementation group C cells remove pyrimidine dimers selectively from the transcribed strand of active genes , 1991, Molecular and cellular biology.

[9]  P. Menichini,et al.  Strand specificity for UV-induced DNA repair and mutations in the Chinese hamster HPRT gene. , 1991, Nucleic acids research.

[10]  J. H. Vos,et al.  Differential introduction of DNA damage and repair in mammalian genes transcribed by RNA polymerases I and II , 1991, Molecular and cellular biology.

[11]  P. van de Putte,et al.  New insights in DNA repair: preferential repair of transcriptionally active DNA. , 1991, Mutagenesis.

[12]  M. Schmid,et al.  Quantitative determination of rDNA transcription units in vertebrate cells. , 1991, Experimental cell research.

[13]  E. Mougey,et al.  News from the nucleolus: rRNA gene expression. , 1991, Trends in biochemical sciences.

[14]  P. Hanawalt,et al.  Selective repair of specific chromatin domains in UV-irradiated cells from xeroderma pigmentosum complementation group C. , 1990, Mutation research.

[15]  L. Thompson,et al.  Cyclobutane-pyrimidine dimer excision in UV-sensitive CHO mutants and the effect of the human ERCC2 repair gene. , 1990, Mutation research.

[16]  R. Schmickel,et al.  Sequence and structure correlation of human ribosomal transcribed spacers. , 1990, Journal of molecular biology.

[17]  L. Mullenders,et al.  The residual repair capacity of xeroderma pigmentosum complementation group C fibroblasts is highly specific for transcriptionally active DNA. , 1990, Nucleic acids research.

[18]  C. Smith,et al.  Clues to the Organization of DNA Repair Systems Gained from Studies of Intragenomic Repair Heterogeneity , 1990 .

[19]  R. Schmickel,et al.  Human ribosomal DNA: novel sequence organization in a 4.5-kb region upstream from the promoter. , 1989, Gene.

[20]  P. Hanawalt,et al.  Induction of the Escherichia coli lactose operon selectively increases repair of its transcribed DNA strand , 1989, Nature.

[21]  J. Sogo,et al.  Two different chromatin structures coexist in ribosomal RNA genes throughout the cell cycle , 1989, Cell.

[22]  P. Hanawalt,et al.  Repair analysis of mitomycin C-induced DNA crosslinking in ribosomal RNA genes in lymphoblastoid cells from Fanconi's anemia patients. , 1989, Mutation research.

[23]  S. A. Leadon,et al.  Differential repair of DNA damage in the human metallothionein gene family , 1988, Molecular and cellular biology.

[24]  J. Wang,et al.  Involvement of DNA topoisomerase I in transcription of human ribosomal RNA genes. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[25]  P. Hanawalt,et al.  Selective removal of transcription-blocking DNA damage from the transcribed strand of the mammalian DHFR gene , 1987, Cell.

[26]  J. Tower,et al.  Transcription of mouse rDNA is regulated by an activated subform of RNA polymerase I , 1987, Cell.

[27]  G. Roeder,et al.  Recombination-stimulating sequences in yeast ribosomal DNA correspond to sequences regulating transcription by RNA polymerase I , 1987, Cell.

[28]  P. Hanawalt,et al.  Preferential DNA repair of an active gene in human cells. , 1986, Proceedings of the National Academy of Sciences of the United States of America.

[29]  P. Hanawalt,et al.  Survival of UV-irradiated mammalian cells correlates with efficient DNA repair in an essential gene. , 1986, Proceedings of the National Academy of Sciences of the United States of America.

[30]  P. Hanawalt,et al.  Differential DNA repair in transcriptionally active and inactive proto-oncogenes: c-abl and c-mos , 1986, Cell.

[31]  M. D'Esposito,et al.  Molecular analysis of the heterogeneity region of the human ribosomal spacer. , 1985, Journal of molecular biology.

[32]  P. Hanawalt,et al.  DNA repair in an active gene: Removal of pyrimidine dimers from the DHFR gene of CHO cells is much more efficient than in the genome overall , 1985, Cell.

[33]  M. Lieberman,et al.  The use of antibodies to 5-bromo-2'-deoxyuridine for the isolation of DNA sequences containing excision-repair sites. , 1984, The Journal of biological chemistry.

[34]  S. Beverley,et al.  Rapid spontaneous dihydrofolate reductase gene amplification shown by fluorescence-activated cell sorting. , 1983, Proceedings of the National Academy of Sciences of the United States of America.

[35]  W. Haseltine,et al.  Quantitation of cyclobutane pyrimidine dimer formation in double- and single-stranded DNA fragments of defined sequence. , 1982, Radiation research.

[36]  J. Erickson,et al.  Structure and variation of human ribosomal DNA: molecular analysis of cloned fragments. , 1981, Gene.

[37]  I B Dawid,et al.  Repeated genes in eukaryotes. , 1980, Annual review of biochemistry.

[38]  N. Arnheim Characterization of mouse ribosomal gene fragments purified by molecular cloning. , 1979, Gene.

[39]  E. Southern,et al.  Heterogeneity of the ribosomal genes in mice and men , 1977, Cell.

[40]  Suraiya Rasheed,et al.  Characterization of a newly derived human sarcoma cell line (HT‐1080) , 1974, Cancer.