Analysis of heterogeneity in Fanconi's anemia patients of different ethnic origin

Zakrzewski, Sabine; Sperling, Karl

doi:10.1007/bf00304547

Cited by 15 publications

(3 citation statements)

References 9 publications

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“…2) normal cells cannot explain this result. By cocultivation of FA and normal cells at a 1:2 ratio, which is much higher than in our case, an increase in chromosomal aberrations in the normal cells and only a slight correction in FA cells were observed (34). The higher growth rate and plating efficiency of MMCR cells seems to account for the data (Fig.…”

Section: Methodssupporting

confidence: 44%

Abnormal response to DNA crosslinking agents of Fanconi anemia fibroblasts can be corrected by transfection with normal human DNA.

Diatloff-Zito¹,

Papadopoulo²,

Averbeck³

et al. 1986

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

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Primary skin fibroblast cell lines from patients with Fanconi anemia were cotransfected with UVirradiated pSV2neo plasmids and high molecular weight DNA from normal human cells. Restoration of a normal cellular resistance to mitomycin C (MMC) was observed provided that a Fanconi anemia cell line is selected for DNA-mediated transformation (neo gene) and that at least two successive rounds of transfection are performed. Cells were selected by taking advantage of the higher proliferation rate and plating efficiency of the MMC resistant transformants. As estimated from reconstruction experiments, the frequency of transfer of MMC resistance lies between 1 and 30 X 10-. The MMC resistance phenotype was maintained for at least 10 generations following transfection. Evidence for DNA-mediated transformation also includes the recovery of a normal pattern of DNA semiconservative synthesis after treatment with 8-methoxypsoralen and 365-nm UV irradiation, and the presence of exogenous pSV2neo DNA sequences was shown by Southern blot analysis. The acquired MMC resistance is probably due to the presence of DNA from normal cells. Indeed, sensitivity to MMC was maintained when Fanconi anemia cells were cotransfected with the UV-irradiated pSV2neo plasmid mixed with their own DNA or with yeast or salmon sperm DNA. These negative results also render unlikely the selection of spontaneous MMC resistant revertants in transfection of Fanconi anemia cells with normal DNA. These experiments establish the prerequisites for the isolation of the gene(s) involved in the response to DNA crosslinking lesions in human cells.Fanconi anemia (FA) is an autosomal recessive disorder in which patients show pancytopenia and predisposition to leukemia and carcinoma (1, 2). These patients show a high level of chromatid-type damage in their lymphocytes and fibroblasts (2-4). Cultured FA cells are unusually sensitive to DNA crosslinking agents by chromosomal criteria (5-9) and by clonogenic cell survival (10-14) whereas their sensitivity to monofunctional agents or to radiation is close to normal (12, 15).A number of antitumor drugs and environmental pollutants produce interstrand crosslinks in DNA. Hence, it is important to understand the consequences of such lesions. Since FA is characterized by an almost specific sensitivity to DNA crosslinking agents it may serve as a model system for research in this area. The nature of the repair defect in FA cells is, however, still debated (11,14,(16)(17)(18) (20) has been shown. However, almost nothing is known about the chromosomal location, expression, and regulation of the genes controlling sensitivity of human cells to DNA crosslinking agents. To investigate these aspects, a project aimed at the isolation of the genes involved was initiated, and we established conditions for the DNA-mediated transfer and expression of wild-type genes in FA cell lines. The isolation and characterization of the transformed population obtained are described here. MATERIALS AND METHODSCell Lines and Cell Cultures. Three...

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Section: Methodssupporting

confidence: 44%

Abnormal response to DNA crosslinking agents of Fanconi anemia fibroblasts can be corrected by transfection with normal human DNA.

Diatloff-Zito¹,

Papadopoulo²,

Averbeck³

et al. 1986

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

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“…Although complementation analyses have demonstrated the presence of at least two distinct FA groups (Zakrzewski and Sperling 1980, Zakrzewski et al . 1983, Duckworth-Rysieki et al 1985, intergenic heterogeneity may not play as extensive a role in this disorder as it does in XP or A-T (Yoshida 1982 (Cockaye 1936(Cockaye , 1946), but is not associated with an increase in cancer incidence .…”

Section: 4 2 Fanconi's Anemia Another Human Disorder Associatedmentioning

confidence: 98%

The Molecular Genetics of the Incision Step in the DNA Excision Repair Process

Rubin

1988

International Journal of Radiation Biology

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This review describes the evolution of research into the genetic basis of how different organisms use the process of excision repair to recognize and remove lesions from their cellular DNA. One particular aspect of excision repair, DNA incision, and how it is controlled at the genetic level in bacteriophage, bacteria, S. cerevisae, D. melanogaster, rodent cells and humans is examined. In phage T4, DNA is incised by a DNA glycosylase-AP endonuclease that is coded for by the denV gene. In E. coli, the products of three genes, uvrA, uvrB and uvrC, are required to form the UVRABC excinuclease that cleaves DNA and releases a fragment 12-13 nucleotides long containing the site of damage. In S. cerevisiae, genes complementing five mutants of the RAD3 epistasis group, rad1, rad2, rad3, rad4 and rad10 have been cloned and analyzed. Rodent cells sensitive to a variety of mutagenic agents and deficient in excision repair are being used in molecular studies to identify and clone human repair genes (e.g. ERCC1) capable of complementing mammalian repair defects. Most studies of the human system, however, have been done with cells isolated from patients suffering from the repair defective, cancer-prone disorder, xeroderma pigmentosum, and these cells are now beginning to be characterized at the molecular level. Studies such as these that provide a greater understanding of the genetic basis of DNA repair should also offer new insights into other cellular processes, including genetic recombination, differentiation, mutagenesis, carcinogenesis and aging.

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“…1982) were the first to show evidence of genetic heterogeneity in FA. Complementation studies in somatic cell hybrids between an SV40-transformed FA cell line and diploid fibroblasts from FA patients suggested the presence of at least two complementation groups (Zakrzewski and Sperling, 1980;1982). DuckworthRysiecki et al (1985) subsequently classified lymphoblastoid FA cell lines into two complementation groups, A and non-A ( = B).…”

Section: Discussionmentioning

confidence: 99%

The Chinese hamster cell mutant V-H4 is homologous to Fanconi anemia (complementation group A)

Arwert

Rooimans²,

Westerveld³

et al. 1991

Cytogenet Genome Res

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V-H4, a mitomycin C (MMC)-sensitive Chinese hamster cell mutant, is phenotypically very similar to Fanconi anemia (FA) cells. Genetic complementation analysis shows that V-H4 belongs to the same complementation group as FA group A cells. Proliferating hybrid cell lines obtained after fusion of V-H4 with normal or FA group B cells show an increased resistance to MMC. Absence of complementation was noted in V-H4 × FA group A hybrid cell lines. This was shown not to be due to the absence of a specific human chromosome. The V-H4 mutant represents the first rodent mutant that is genotypically similar to FA complementation group A cells.

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Analysis of heterogeneity in Fanconi's anemia patients of different ethnic origin

Cited by 15 publications

References 9 publications

Abnormal response to DNA crosslinking agents of Fanconi anemia fibroblasts can be corrected by transfection with normal human DNA.

Abnormal response to DNA crosslinking agents of Fanconi anemia fibroblasts can be corrected by transfection with normal human DNA.

The Molecular Genetics of the Incision Step in the DNA Excision Repair Process

The Chinese hamster cell mutant V-H4 is homologous to Fanconi anemia (complementation group A)

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