[1] Techniques of localization of proteins and nucleoproteins in the cell nucleus by high resolution autoradiography and cytochemistry

Bouteille, M.; Dupuy-Coin, A.M.; Moyne, Gilles

doi:10.1016/s0076-6879(75)40003-9

Cited by 22 publications

(7 citation statements)

References 45 publications

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“…First, they show a more general distribution of nucleic acids in biological materials. The distribution and density of uridinelthymidine-labeled molecules detected by autoradiography depends on the duration of the labeling procedure and on any postincubation or chase experiments that might be performed (Bouteille et al, 1975;Fakan, 1976). Likewise, the labeling patterns obtained with enzyme-gold complexes differ according to the enzyme used (Cheniclet and Bendayan, 1990).…”

Section: Potential Value and Limitationsmentioning

confidence: 99%

Nucleic acid compartmentalization within the cell nucleus by in situ transferase‐immunogold techniques

Thiry

1995

Microscopy Res & Technique

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In the present review, we report on recent results obtained by in situ transferase-immunogold techniques as to the ultrastructural distribution of DNA and RNA within the cell nucleus. Special emphasis is placed on the various nucleolar components and the various enigmatic structures of the extranucleolar region: interchromatin granules, coiled bodies, and simple nuclear bodies. These data are discussed in the light of our current understanding of the functional organization of the cell nucleus.

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Section: Potential Value and Limitationsmentioning

confidence: 99%

Nucleic acid compartmentalization within the cell nucleus by in situ transferase‐immunogold techniques

Thiry

1995

Microscopy Res & Technique

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“…The Chinese hamster cell line DC-3F and the 9-hydroxyellipticine-resistant subline DC-3F/ 9-OH-E have been described (25), as have the media and growth conditions (25,26 was immediately withdrawn. The cells were prepared for autoradiography as described (27,28).…”

mentioning

confidence: 99%

Suramin is an inhibitor of DNA topoisomerase II in vitro and in Chinese hamster fibrosarcoma cells.

Bojanowski

Lelièvre

Markovits

et al. 1992

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

107

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The antitrypanosomal and antifiliarial drug suramin is currently under investigation for treatment of advanced malignancies including prostatic cancer, adrenocortical cancer, and some lymphomas and sarcomas. Here we show that suramin is a potent inhibitor of the nuclear enzyme DNA topoisomerase II. Suramin inhibited purified yeast topoisomerase II with an IC5s of about 5 FM, as measured by decatenation or relaxation assays. Suramin did not stabilize the covalent DNA-topoisomerase II reaction intermediate ("cleavable complex"), whereas other inhibitors of this enzyme, such as amsacrine, etoposide, and the ellipticines, are known to stabilize the intermediate. In contrast, the presence of suramin strongly inhibited the cleavable-complex formation induced by amsacrine or etoposide. Accumulation of the endogenous cleavable complex was also inhibited. Suramin entered the nucleus of DC-3F Chinese hamster fibrosarcoma cells exposed to radiolabeled suramin for 24 hr as shown by both optic and electron microscopy. The suramin present in the nucleus seemed to interact with topoisomerase II, since suramin reduced the number of amsacrine-induced protein-associated DNA strand breaks in DC-3F cells and protected these cells from the cytotoxic action of amsacrine. Cells resistant to 9-hydroxyellipticine, which have been shown to have an altered topoisomerase II activity, are about 7-fold more resistant to suramin than the sensitive parental cells as shown by 72-hr growth inhibition assay. Our results suggest that DNA topoisomerase II is a target of suramin action and that this action may play a role in the cytotoxic activity of suramin.Suramin is a hexasulfated naphthylurea that has been used in the treatment of trypanosomiasis (sleeping sickness) and onchocerciasis for more than half a century (1). Mor recently, suramin was shown to prevent infection of T lymphocytes by human immunodeficiency virus in vitro (2). This led to clinical trials of the compound in AIDS patients, where it showed little therapeutic activity (3, 4). However, during these trials a complete clinical response was noted in a patient with Kaposi sarcoma and non-Hodgkin lymphoma (4), and subsequent studies showed that suramin was active in the treatment of several metastatic cancers such as renal cancer, adrenocarcinoma, lymphoma, and prostate cancer (5, 6).Suramin inhibits the binding of platelet-derived growth factor, epidermal growth factor, basic fibroblast growth factor, transforming growth factor /3, and insulin-like growth factor to their specific cell surface receptors (7-11). The drug also affects the activity of protein kinase C and inhibits phosphatidylinositol kinase and diacylglycerol kinase (12, 13). Several nuclear enzymes are also inhibited by suramin. These include DNA and RNA polymerases, terminal deoxynucleotidyltransferase, and reverse transcriptase (14-17).Suramin can induce cell differentiation in several different systems (18,19) and can inhibit tumor cell invasion (20).Here we demonstrate that suramin is a potent inhibitor of...

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“…Compared to all the other techniques used so far to detect DNA, the in situ TdT method also shows a more general distribution of nucleic acids in biological materials. The autoradiographic techniques allow detection of only a fraction of the DNA, that which was freshly synthetized by replication during a certain period of time (Bouteille et al 1975;Fakan 1976Fakan , 1978. Likewise, the DNA configuration affects the labeling pattern obtained with DNase I-gold complexes (Bendayan 1981a(Bendayan , b, 1984.…”

Section: In Situ Terminal Deoxynucleotidyl Transferase-immunogold Metmentioning

confidence: 99%

“…Furthermore, the 5' end of single-stranded DNA present at the surface of ultrathin sections can, in principle, be labeled. Unlike autoradiography after uptake of labeled ribonucleotides (Bouteille et al 1975;Fakan 1976Fakan , 1978, the in situ-in vitro transcription technique avoids the danger of displacement of newly synthetized molecules from the site of synthesis to the site of accumulation. With the latter approach, however, as with the in situ nick translation-immunogold technique (Thiry 1991a, b), it is not possible to distinguish transcriptionally active from potentially active regions.…”

Section: Active Dna Region Identified By the In Situ-in Vitro Transcrmentioning

confidence: 99%

“…The distribution and density of uridine-labeled molecules detected by autoradiography depends strictly on the duration of the labeling procedure and on any postincubation or chase experiments that may be performed (Bouteille et al 1975;Fakan 1976Fakan , 1978. Likewise, the labeling patterns obtained with RNase-gold complexes depend on the nature of the enzyme used (Cheniclet and Bendayan 1990).…”

Section: Active Dna Region Identified By the In Situ-in Vitro Transcrmentioning

confidence: 99%

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New approaches to in situ detection of nucleic acids

Thiry

1995

Histochem Cell Biol

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The present paper reviews recent results obtained by different molecular biology-based, immunocytological approaches to the localization and identification of nucleic acids in sections of biological material. Examples of sensitive, high-resolution detection methods for RNA, DNA or specialized DNA regions are presented. Special emphasis is placed on the potential values and limitations of these new methods.

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[1] Techniques of localization of proteins and nucleoproteins in the cell nucleus by high resolution autoradiography and cytochemistry

Cited by 22 publications

References 45 publications

Nucleic acid compartmentalization within the cell nucleus by in situ transferase‐immunogold techniques

Nucleic acid compartmentalization within the cell nucleus by in situ transferase‐immunogold techniques

Suramin is an inhibitor of DNA topoisomerase II in vitro and in Chinese hamster fibrosarcoma cells.

New approaches to in situ detection of nucleic acids

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