The Effect of Ultraviolet Light on Arrested Human Diploid Cell Populations

Kantor, George J.; Warner, Cecilia A.; Hull, D.R.

doi:10.1111/j.1751-1097.1977.tb09174.x

Cited by 51 publications

(18 citation statements)

References 24 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…For comparison, we included an analysis of proliferating and quiescent 1BR.3. In complete medium, 1BR.3 proliferate rapidly but, when maintained under low serum conditions for 7 days, these cells enter stationary phase and arrest in G1 (Kantor et al, 1977). Figure 5 shows the abundance of each mRNA using isoform-specific primers for qPCR.…”

Section: Characterization Of Fancd2 Expression In Sh-sy5ymentioning

confidence: 99%

Alcohol Induces DNA Damage and the Fanconi Anemia D2 Protein Implicating FANCD2 in the DNA Damage Response Pathways in Brain

Rulten

Hodder

Ripley

et al. 2008

Alcoholism Clin & Exp Res

View full text Add to dashboard Cite

In contrast to other DNA damaging agents, ethanol/acetaldehyde generated DNA strand breaks without inducing ubiquitination of FANCD2, despite increasing protein levels in the nucleus. These data are consistent with recent reports that suggest the Fanconi anemia pathway plays an important role in the adult brain in response to DNA damage. Further work is required to establish what this role is, in particular the potential function of nonubiquitinated FANCD2 and its role in the DNA damage response in postmitotic neurons and neural precursor cells.

show abstract

Section: Characterization Of Fancd2 Expression In Sh-sy5ymentioning

confidence: 99%

Alcohol Induces DNA Damage and the Fanconi Anemia D2 Protein Implicating FANCD2 in the DNA Damage Response Pathways in Brain

Rulten

Hodder

Ripley

et al. 2008

Alcoholism Clin & Exp Res

View full text Add to dashboard Cite

show abstract

“…Nondividing (arrested) populations of human fibroblasts were established and maintained by a modification of established procedures (18,19). Freshly trypsinized fibroblasts were plated into a series of 6-cm (diameter) tissue culture dishes at a cell density of 1.6 x 105 cells per dish and were incubated for 24 hr in medium supplemented with 15% serum.…”

Section: Methodsmentioning

confidence: 99%

Exposure of nondividing populations of primary human fibroblasts to UV (254 nm) radiation induces a transient enhancement in capacity to repair potentially lethal cellular damage.

Tyrrell¹

1984

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

Nondividing (arrested) populations of primary human fibroblasts from normal individuals exposed to an initial dose (1.5 or 3 J m-2) of far-UV (254 nm) radiation and then incubated in medium containing low (0.5%) serum develop enhanced resistance to inactivation of cloning efficiency by a second (challenge) dose of UV. The resistance develops within 2-4 days, after which there is a decline. Resistance develops to a higher degree and more rapidly (1-2 days) in cells derived from patients with the variant form of xeroderma pigmentosum. Excision-deficient cells from xeroderma pigmentosum complementation group A individuals also develop UV resistance after a lower (0.2 J m-2) exposure to UV. Enhanced UV resistance does not develop in UV-irradiated cell populations incubated with the protein synthesis inhibitor cycloheximide (5 IPM). These observations are consistent with the interpretation that exposure of human fibroblasts to low doses of UV induces synthesis of a protein involved in a metabolic pathway that transiently enhances the capacity of cells to repair potentially lethal damage resulting from a subsequent dose of UV.The genetic control and biochemistry of an error-prone far-UV inducible repair system in Escherichia coli has been extensively studied (see ref. 1 for overview). Recent experiments have begun to clarify the nature of the inducing signal (2) and enzymology (3) of the process. The existence of a similar phenomenon in eukaryotic cells has been largely inferred from viral reactivation studies (4). The survival of many types of UV-damaged virus is enhanced by pre-exposure of the appropriate host cells to UV radiation and incubation for an appropriate time prior to viral infection (see ref.5 for review). The nature and degree of error-proneness of the response, as deduced from enhanced mutagenesis of infecting virus (6, 7), appears to vary considerably with the particular virus and host cell system employed. For example, the mutability of intact but not UV-irradiated parvovirus H1 is enhanced in UV pretreated human or rat cells (8), whereas only enhanced mutability of UV-treated virus is seen when simian virus 40 infects UV-treated monkey cells (7). Findings opposite to the latter have been reported (9, 10).Biochemical measurements of UV-induced responses in mammalian cells have been controversial. A report of enhancement of post-replication repair by a low-dose UV-conditioning exposure (11) may also be interpreted (12) on the basis of the abnormal state of DNA replication at the time of the second exposure. Nevertheless, UV exposure of human fibroblasts has been shown to enhance levels of plasminogen activator (13) and DNA ligase (14), and agents that arrest DNA replication have been shown recently to induce synthesis of a nuclear membrane-bound protein in cell lines derived from B lymphocytes (15).Little is known about whether UV treatment actually enhances the capacity of mammalian cells to survive subsequent challenge doses of UV. Split-dose experiments are often difficult to interpret (16)...

show abstract

“…It has been shown that diploid human cells that have been arrested by reducing the serum concentration to 0.5% have normal levels of UDS (8). Incubation of various XP cell lines in 0.5% serum for a minimum of 14 days usually reduces the amount of replication synthesis during a 2-hr labeling period to <0.1%.…”

Section: Resultsmentioning

confidence: 99%

Transient complementation of xeroderma pigmentosum cells by microinjection of poly(A)+ RNA.

Legerski¹,

Brown²,

Peterson³

et al. 1984

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

An assay has been developed in which excision repair deficiency of xeroderma pigmentosum cells is transiently complemented, as measured by unscheduled DNA synthesis, by microinjection of cytoplasmic poly(A)+ RNA derived from HeLa cells. Four different complementation groups of xeroderma pigmentosum have been assayed. Groups A and G showed complementation, whereas groups D and F did not. Survival for cells in each of the groups subsequent to microinjection was =75%. Approximately 10-25% of surviving cells from groups A and G were complemented, as judged by nearnormal unscheduled DNA synthesis. Fractionation of cytoplasnic poly(A)+ RNA on a 15-30% nondenaturing sucrose gradient and subsequent microinjection of the individual fractions indicate that repair mRNAs that complement xerbderma pigmentosum groups A and G sediment at approximately 11 S and 12 S, respectively. This assay should be of great utility in the cloning and biochemical analysis of DNA repair genes.Xeroderma pigmentosum (XP) is an autosomal, recessive disease of humans characterized by dermatological and ocular symptoms and a high predisposition to skin cancer after exposure to sunlight. Some forms of XP have associated neurological disorders (1) and possible immunological abnormalities. The primary biochemical defect in most XP cells is thought to be an inability to initiate excision repair synthesis of their DNA after exposure to UV irradiation (2, 3). Recently, the feasibility of transiently complementing mutant cell lines by direct microinjection of unenriched cytoplasmic poly(A)+ RNA has been demonstrated (4, 5). In this report, we show that microinjection of cytoplasmic poly(A)+ RNA derived from HeLa cells can transiently complement XP cells, as determined by a temporary increase in the level of unscheduled DNA synthesis (UDS). In addition, fractionation of poly(A)+ RNA on nondenaturing sucrose gradients and subsequent microinjection of the individual fractions indicated that the repair mRNAs that complement XP-A and -G cells sedimented at unique positions. MATERIALS AND METHODSCell Cultures. Fibroblast cell strains XP25RO (XP-A), XP3NE (XP-D), XP2YO (XP-F), and XP2BI (XP-G) were obtained from the Human Genetic Mutant Cell Repository. The cells were routinely grown in minimal essential medium (Flow Laboratories) containing 20% fetal calf serum (Irvine Scientific), penicillin G (100 units/ml), streptomycin (100 ,ug/ml), and glutamine (2 mM). HeLa S3 cells were grown in suspension culture containing minimal essential medium and 10% calf serum (Flow Laboratories).Isolation of Cytoplasmic Poly(A)+ RNA. The procedure for preparation of poly(A)+ RNA was a modification of the method described by Lin et al. (5). Approximately 5 x 109 cells were pelleted, washed twice with saline, and resuspended in 50 ml of ice-cold lysis buffer containing 30 mM Tris-HCl (pH 7.4), 30 mM KCI, 0.1 M NaCi, heparin (1 mg/ml), 0.15% Nonidet P-40, and 10 mM vanadyl ribonucleoside complex (Bethesda Research Laboratories). After 10 min on ice the sample was centrifuge...

show abstract

The Effect of Ultraviolet Light on Arrested Human Diploid Cell Populations

Cited by 51 publications

References 24 publications

Alcohol Induces DNA Damage and the Fanconi Anemia D2 Protein Implicating FANCD2 in the DNA Damage Response Pathways in Brain

Alcohol Induces DNA Damage and the Fanconi Anemia D2 Protein Implicating FANCD2 in the DNA Damage Response Pathways in Brain

Exposure of nondividing populations of primary human fibroblasts to UV (254 nm) radiation induces a transient enhancement in capacity to repair potentially lethal cellular damage.

Transient complementation of xeroderma pigmentosum cells by microinjection of poly(A)+ RNA.

Contact Info

Product

Resources

About