Effect of UV light on small nuclear RNA synthesis: increased inhibition during postirradiation cell incubation

1986 ◽  
Vol 6 (3) ◽  
pp. 745-750
Author(s):  
D S Morra ◽  
B P Eliceiri ◽  
G L Eliceiri
Keyword(s):  
Uv Light ◽  
Human Fibroblasts ◽  
Complementation Group ◽  
Small Nuclear Rna ◽  
Dna Breaks ◽  
Single Stranded Dna ◽  
Inactivation Curve ◽  
Torsional Strain ◽  
Pyrimidine Dimers ◽  
Cell Incubation

It has been shown previously that the synthesis of small nuclear RNAs (snRNAs) U1, U2, U3, U4, and U5, in contrast to that of all other RNA species tested, decreases markedly within 2 h of cell incubation after exposure to UV light (254 nm), while pyrimidine dimers are being removed from DNA. We examined the possibility that the postirradiation cell incubation-dependent, UV light-induced inhibition of snRNA synthesis might reflect hypersensitivity of the snRNA transcriptional domains to single-stranded DNA nicks or relaxation of DNA torsional stress or both that occur during DNA repair. This late suppression of snRNA biosynthesis was as pronounced in UV light-irradiated (DNA incision-deficient) xeroderma pigmentosum fibroblasts (complementation group A) as in irradiated normal human fibroblasts. The synthesis of snRNAs was not preferentially sensitive to gamma radiation (which produces single-stranded DNA breaks) or novobiocin or nalidixic acid (which induce DNA relaxation). Neither of these two drugs prevented the UV light-induced inhibition of snRNA synthesis observed during postirradiation cell incubation. These results suggest that the late suppression of snRNA synthesis does not result from hypersensitivity of snRNA transcriptional domains to single-stranded DNA cleavages or relaxation of DNA torsional strain. The UV light-induced late inhibition of snRNA synthesis: shows an inactivation curve whose slope differs from that observed immediately after irradiation; is seen in untransformed cells as well as established cells lines; and has been conserved between birds and mammals.

scholarly journals Effect of UV light on small nuclear RNA synthesis: increased inhibition during postirradiation cell incubation.

1986 ◽  
Vol 6 (3) ◽  
pp. 745-750 ◽  
Author(s):  
D S Morra ◽  
B P Eliceiri ◽  
G L Eliceiri
Keyword(s):  
Uv Light ◽  
Human Fibroblasts ◽  
Complementation Group ◽  
Small Nuclear Rna ◽  
Dna Breaks ◽  
Single Stranded Dna ◽  
Inactivation Curve ◽  
Torsional Strain ◽  
Pyrimidine Dimers ◽  
Cell Incubation

It has been shown previously that the synthesis of small nuclear RNAs (snRNAs) U1, U2, U3, U4, and U5, in contrast to that of all other RNA species tested, decreases markedly within 2 h of cell incubation after exposure to UV light (254 nm), while pyrimidine dimers are being removed from DNA. We examined the possibility that the postirradiation cell incubation-dependent, UV light-induced inhibition of snRNA synthesis might reflect hypersensitivity of the snRNA transcriptional domains to single-stranded DNA nicks or relaxation of DNA torsional stress or both that occur during DNA repair. This late suppression of snRNA biosynthesis was as pronounced in UV light-irradiated (DNA incision-deficient) xeroderma pigmentosum fibroblasts (complementation group A) as in irradiated normal human fibroblasts. The synthesis of snRNAs was not preferentially sensitive to gamma radiation (which produces single-stranded DNA breaks) or novobiocin or nalidixic acid (which induce DNA relaxation). Neither of these two drugs prevented the UV light-induced inhibition of snRNA synthesis observed during postirradiation cell incubation. These results suggest that the late suppression of snRNA synthesis does not result from hypersensitivity of snRNA transcriptional domains to single-stranded DNA cleavages or relaxation of DNA torsional strain. The UV light-induced late inhibition of snRNA synthesis: shows an inactivation curve whose slope differs from that observed immediately after irradiation; is seen in untransformed cells as well as established cells lines; and has been conserved between birds and mammals.

Cell lines from xeroderma pigmentosum complementation group A lack a single-stranded-DNA-binding activity

1983 ◽  
Vol 3 (7) ◽  
pp. 667-674 ◽  
Author(s):  
Urs Kuhnlein ◽  
Siu Sing Tsang ◽  
Opal Lokken ◽  
Silvian Tong ◽  
Daniel Twa
Keyword(s):  
Dna Binding ◽  
Cell Lines ◽  
Xeroderma Pigmentosum ◽  
Human Fibroblasts ◽  
Complementation Group ◽  
Binding Activity ◽  
Single Stranded Dna ◽  
Dna Binding Activity ◽  
A Cell ◽  
D Cell

Human fibroblasts and HeLa cells contain two major DNA-binding activities for superhelical DNA, which can be separated by phosphocellulose chromatography. The DNA-binding activity which elutes first from the column coelutes with and is probably identical to a single-stranded-DNA-binding activity. The second activity has been characterized previously. It binds preferentially to super-helical DNA containing DNA damage, but does not bind to single-stranded DNA. Five cell lines derived from patients with the repairdeficiency syndrome xeroderma pigmentosum (XP) were analyzed for the presence of these binding activities. Four of the cell lines were from the A-complementation group and one was from the D-complementation group of XP. The binding activity with preference for damaged DNA was present in all cell lines. The single-stranded-DNA-binding activity was present in the XP-D cell line but was absent or reduced in all of the four XP-A cell lines tested.

Mechanisms of tolerance to DNA lesions in mammalian cells

1981 ◽  
Vol 14 (3) ◽  
pp. 381-432 ◽  
Author(s):  
Rogerio Meneghini ◽  
Carlos F. M. Menck ◽  
R. Ivan Schumacher
Keyword(s):  
Genetic Disease ◽  
Mammalian Cells ◽  
Uv Light ◽  
Human Fibroblasts ◽  
Dna Lesions ◽  
Genetic Integrity ◽  
Dna Lesion ◽  
Repair Enzymes ◽  
Pyrimidine Dimers ◽  
A Cell

In recent years it has become clear that different pathways are involved in the process of removing lesions from DNA. In spite of a continuous surveillance of the genetic integrity by repair enzymes, quite often lesions are not eliminated before the portion of the genome where they have been inserted is used for DNA replication or transcription. Actually, the number of unexcised lesions a cell can tolerate without significantly losing its capacity to reproduce is surprising. As an example, human fibroblasts from certain patients with the genetic disease xeroderma pigmentosum (XP)† are virtually unable to excise pyrimidine dimers, the major DNA lesion produced by short-wavelength UV light.

A Requirement for Recombinational Repair in Saccharomyces cerevisiae Is Caused by DNA Replication Defects of mec1 Mutants

Genetics ◽  
1999 ◽  
Vol 153 (2) ◽  
pp. 595-605 ◽  
Author(s):  
Bradley J Merrill ◽  
Connie Holm
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Dna Replication ◽  
Dna Synthesis ◽  
Synthetic Lethality ◽  
Velocity Sedimentation ◽  
Recombinational Repair ◽  
Repair Pathway ◽  
Dna Breaks ◽  
Single Stranded Dna ◽  
Double Stranded Dna

Abstract To examine the role of the RAD52 recombinational repair pathway in compensating for DNA replication defects in Saccharomyces cerevisiae, we performed a genetic screen to identify mutants that require Rad52p for viability. We isolated 10 mec1 mutations that display synthetic lethality with rad52. These mutations (designated mec1-srf for synthetic lethality with rad-fifty-two) simultaneously cause two types of phenotypes: defects in the checkpoint function of Mec1p and defects in the essential function of Mec1p. Velocity sedimentation in alkaline sucrose gradients revealed that mec1-srf mutants accumulate small single-stranded DNA synthesis intermediates, suggesting that Mec1p is required for the normal progression of DNA synthesis. sml1 suppressor mutations suppress both the accumulation of DNA synthesis intermediates and the requirement for Rad52p in mec1-srf mutants, but they do not suppress the checkpoint defect in mec1-srf mutants. Thus, it appears to be the DNA replication defects in mec1-srf mutants that cause the requirement for Rad52p. By using hydroxyurea to introduce similar DNA replication defects, we found that single-stranded DNA breaks frequently lead to double-stranded DNA breaks that are not rapidly repaired in rad52 mutants. Taken together, these data suggest that the RAD52 recombinational repair pathway is required to prevent or repair double-stranded DNA breaks caused by defective DNA replication in mec1-srf mutants.

scholarly journals The effect of flanking bases on direct and triplet sensitized cyclobutane pyrimidine dimer formation in DNA depends on the dipyrimidine, wavelength and the photosensitizer

2021 ◽  
Author(s):  
Chen Lu ◽  
Natalia Eugenia Gutierrez-Bayona ◽  
John-Stephen Taylor
Keyword(s):  
Uv Light ◽  
Pyrimidine Dimer ◽  
Flanking Sequence ◽  
Quenching Mechanism ◽  
Cyclobutane Pyrimidine Dimers ◽  
Sequence Dependence ◽  
Pyrimidine Dimers ◽  
Skin Cancers ◽  
A Charge ◽  
C To T Mutations

Abstract Cyclobutane pyrimidine dimers (CPDs) are the major products of DNA produced by direct absorption of UV light, and result in C to T mutations linked to human skin cancers. Most recently a new pathway to CPDs in melanocytes has been discovered that has been proposed to arise from a chemisensitized pathway involving a triplet sensitizer that increases mutagenesis by increasing the percentage of C-containing CPDs. To investigate how triplet sensitization may differ from direct UV irradiation, CPD formation was quantified in a 129-mer DNA designed to contain all 64 possible NYYN sequences. CPD formation with UVB light varied about 2-fold between dipyrimidines and 12-fold with flanking sequence and was most frequent at YYYR and least frequent for GYYN sites in accord with a charge transfer quenching mechanism. In contrast, photosensitized CPD formation greatly favored TT over C-containing sites, more so for norfloxacin (NFX) than acetone, in accord with their differing triplet energies. While the sequence dependence for photosensitized TT CPD formation was similar to UVB light, there were significant differences, especially between NFX and acetone that could be largely explained by the ability of NFX to intercalate into DNA.

scholarly journals Base excision repair causes age-dependent accumulation of single-stranded DNA breaks that contribute to Parkinson disease pathology

Cell Reports ◽  
2021 ◽  
Vol 36 (10) ◽  
pp. 109668
Author(s):  
Tanima SenGupta ◽  
Konstantinos Palikaras ◽  
Ying Q. Esbensen ◽  
Georgios Konstantinidis ◽  
Francisco Jose Naranjo Galindo ◽  
...  
Keyword(s):  
Parkinson Disease ◽  
Base Excision Repair ◽  
Excision Repair ◽  
Dna Breaks ◽  
Single Stranded Dna ◽  
Age Dependent ◽  
Base Excision

scholarly journals Single-Stranded DNA Breaks Adjacent to Cytosines Occur during Ig Gene Class Switch Recombination

2004 ◽  
Vol 173 (5) ◽  
pp. 3223-3229 ◽  
Author(s):  
Arulvathani Arudchandran ◽  
Ralph M. Bernstein ◽  
Edward E. Max
Keyword(s):  
Class Switch Recombination ◽  
Dna Breaks ◽  
Single Stranded Dna ◽  
Class Switch

Xeroderma pigmentosum complementation group C cells remove pyrimidine dimers selectively from the transcribed strand of active genes

1991 ◽  
Vol 11 (8) ◽  
pp. 4128-4134
Author(s):  
J Venema ◽  
A van Hoffen ◽  
V Karcagi ◽  
A T Natarajan ◽  
A A van Zeeland ◽  
...  
Keyword(s):  
Xeroderma Pigmentosum ◽  
Mammalian Cells ◽  
Excision Repair ◽  
Complementation Group ◽  
C Cells ◽  
Dhfr Gene ◽  
Normal Cells ◽  
Pyrimidine Dimers ◽  
Normal Human ◽  
Active Genes

We have measured the removal of UV-induced pyrimidine dimers from DNA fragments of the adenosine deaminase (ADA) and dihydrofolate reductase (DHFR) genes in primary normal human and xeroderma pigmentosum complementation group C (XP-C) cells. Using strand-specific probes, we show that in normal cells, preferential repair of the 5' part of the ADA gene is due to the rapid and efficient repair of the transcribed strand. Within 8 h after irradiation with UV at 10 J m-2, 70% of the pyrimidine dimers in this strand are removed. The nontranscribed strand is repaired at a much slower rate, with 30% dimers removed after 8 h. Repair of the transcribed strand in XP-C cells occurs at a rate indistinguishable from that in normal cells, but the nontranscribed strand is not repaired significantly in these cells. Similar results were obtained for the DHFR gene. In the 3' part of the ADA gene, however, both normal and XP-C cells perform fast and efficient repair of either strand, which is likely to be caused by the presence of transcription units on both strands. The factor defective in XP-C cells is apparently involved in the processing of DNA damage in inactive parts of the genome, including nontranscribed strands of active genes. These findings have important implications for the understanding of the mechanism of UV-induced excision repair and mutagenesis in mammalian cells.

Induction of metallothionein and other mRNA species by carcinogens and tumor promoters in primary human skin fibroblasts

1986 ◽  
Vol 6 (5) ◽  
pp. 1760-1766
Author(s):  
P Angel ◽  
A Pöting ◽  
U Mallick ◽  
H J Rahmsdorf ◽  
M Schorpp ◽  
...  
Keyword(s):  
Uv Light ◽  
Human Fibroblasts ◽  
Nucleic Acid Hybridization ◽  
Tumor Promoter ◽  
Cdna Clones ◽  
Tumor Promoters ◽  
Human Skin Fibroblasts ◽  
Cdna Sequencing ◽  
Transcript Levels ◽  
Inducible Protein

We used nucleic acid hybridization and cDNA cloning techniques to isolate human sequences that respond to the tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA). These clones were used as probes to examine changes of gene expression that occurred after the proliferation of exponentially growing primary human fibroblasts was arrested. Transcript levels detected by these probes were increased coordinately by treatment of the cells with UV light, mitomycin C, TPA, or the UV light-induced extracellular protein synthesis-inducing factor EPIF (M. Schorpp, U. Mallick, H. J. Rahmsdorf, and P. Herrlich, Cell 37:861-868, 1984). Proteins coded for by these transcripts were characterized by hybrid-promoted translation and by cDNA sequencing. One of the cDNA clones was homologous to the metallothionein IIa gene, and one set of related clones selected RNA for the secreted TPA-inducible protein XHF1 (U. Mallick, H. J. Rahmsdorf, N. Yamamoto, H. Ponta, R.-D. Wegner, and P. Herrlich, Proc. Natl. Acad. Sci. USA 79:7886-7890, 1982).

The RAD7 and RAD16 genes, which are essential for pyrimidine dimer removal from the silent mating type loci, are also required for repair of the nontranscribed strand of an active gene in Saccharomyces cerevisiae

1994 ◽  
Vol 14 (9) ◽  
pp. 6135-6142
Author(s):  
R Verhage ◽  
A M Zeeman ◽  
N de Groot ◽  
F Gleig ◽  
D D Bang ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Mating Type ◽  
Excision Repair ◽  
Complementation Group ◽  
Pyrimidine Dimer ◽  
Gene Products ◽  
Pyrimidine Dimers ◽  
Nucleotide Excision ◽  
Active Genes ◽  
Silent Mating Type Loci

The rad16 mutant of Saccharomyces cerevisiae was previously shown to be impaired in removal of UV-induced pyrimidine dimers from the silent mating-type loci (D. D. Bang, R. A. Verhage, N. Goosen, J. Brouwer, and P. van de Putte, Nucleic Acids Res. 20:3925-3931, 1992). Here we show that rad7 as well as rad7 rad16 double mutants have the same repair phenotype, indicating that the RAD7 and RAD16 gene products might operate in the same nucleotide excision repair subpathway. Dimer removal from the genome overall is essentially incomplete in these mutants, leaving about 20 to 30% of the DNA unrepaired. Repair analysis of the transcribed RPB2 gene shows that the nontranscribed strand is not repaired at all in rad7 and rad16 mutants, whereas the transcribed strand is repaired in these mutants at a fast rate similar to that in RAD+ cells. When the results obtained with the RPB2 gene can be generalized, the RAD7 and RAD16 proteins not only are essential for repair of silenced regions but also function in repair of nontranscribed strands of active genes in S. cerevisiae. The phenotype of rad7 and rad16 mutants closely resembles that of human xeroderma pigmentosum complementation group C (XP-C) cells, suggesting that RAD7 and RAD16 in S. cerevisiae function in the same pathway as the XPC gene in human cells. RAD4, which on the basis of sequence homology has been proposed to be the yeast XPC counterpart, seems to be involved in repair of both inactive and active yeast DNA, challenging the hypothesis that RAD4 and XPC are functional homologs.

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