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J Filipski

Publications and source records attributed to J Filipski.

At least 37 records · Page 2Linked to original sources
PubMed1987

Correlation between molecular clock ticking, codon usage fidelity of DNA repair, chromosome banding and chromatin compactness in germline cells.

The vertebrate genome is built of long DNA regions, relatively homogeneous in GC content, which likely correspond to bands on stained chromosomes. Large differences in composition have been found among DNA regions belonging to the same genome. They are paralleled by differences in codon usage in genes differently localized. The hypothesis presented here asserts that these differences in composition are caused by different mutational bias of alpha and beta DNA polymerases, these polymerases being involved to different extents in the repair of DNA lesions in compact and relaxed chromatin, respectively, in germline cells.

Animals↗
PubMed1987

Nonrandom distribution of MMTV proviral sequences in the mouse genome.

Integrated sequences of mouse mammary tumor virus (MMTV) have been localized in the genomes of five inbred mouse strains (Balb/c, C3H, DBA/2, A.TH, 129-SV) and one mammary tumor cell line (GR). Two major classes of MMTV sequences have been detected in mouse DNA fractions as obtained by Cs2SO4/BAMD (3,6-bis-(acetatomercurimethyl)dioxane) density gradient centrifugation. The first one corresponds to previously described endogenous sequences (Mtv loci), whereas the second one corresponds to endogenous sequences not previously known, and/or recently acquired; in the case of GR cells exogenous sequences may also be present in this class. The genome distribution is somewhat different for the two classes of sequences, the first one being practically only present in the lightest DNA segments of the mouse genome (GC congruent to 38%); the second one being also represented in heavier segments (GC congruent to 43%). This integration pattern suggests that "ancient" endogenous sequences are practically only localized in genome segments of roughly matching composition, whereas exogenous and recently acquired endogenous MMTV sequences may also be present in heavier fractions.

Animals↗
PubMed1987

Two distinct compositional classes of vertebrate gene-bearing DNA stretches, their structures and possible evolutionary origin.

Genomes of vertebrates are built of long, compositionally uniform DNA regions differing in guanine and cytidine (G + C) content. Examination of G + C distribution and CpG dinucleotide frequency in the longest stretches of vertebrate DNA base sequences available show that the long-range structural features are correlated with the structure of genes. Two classes of DNA stretches are conspicuous: (i) the stretches having low G + C content and low CpG doublet frequency and (ii) stretches rich in G + C containing CpG-rich islands. Both classes show other compositional islands containing exons. These structural features result from evolutionary pressures acting on the DNA or RNA level, as well as from mutations and repair differently biased in different genomic compartments. The analysis presented provides a rationale for a discussion of evolution of the long-range structural characteristics of DNA.

Animals↗
PubMed1986

Genomic localization of hepatitis B virus in a human hepatoma cell line.

The integration of hepatitis B viral sequences in the human hepatoma Alexander cell line has been investigated after fractionation of the cell line DNA by centrifugation in a Cs2SO4/BAMD (3,6-(bis-acetato mercurimethyl) dioxane) density gradient. Eight out of nine integrated viral sequences were localized in DNA component H3, which only represents 4% of the human genome and matches the base composition of HBV sequences. These results indicate a targeting and/or a higher stability of the latter in a specific, small compartment of the host genome.

Carcinoma, Hepatocellular↗
PubMed1986

Gene distribution and nucleotide sequence organization in the mouse genome.

Mouse DNA was fractionated by preparative centrifugation in density gradients of Cs2SO4 containing 3,6-bis(acetatomercurimethyl)dioxane (BAMD). The effects of temperature, BAMD/nucleotide molar ratio and solvent on the fractionation were explored. The fractions so obtained were investigated by analytical centrifugation in CsCl density gradient and by hybridization with a number of gene probes. These approaches led to the definition of satisfactory conditions for the rapid fractionation of mouse DNA; to the localization of a number of genes in mouse DNA fractions; and to a better understanding of the mosaic organization of the mouse genome and, more specifically, to a better estimate of both the intermolecular and intramolecular compositional heterogeneity of mouse DNA in the (75-150) X 10(3)-base size range.

Actins↗
PubMed1986

Gene distribution and nucleotide sequence organization in the human genome.

Human DNA was fractionated by centrifugation in Cs2SO4 density gradients containing 3,6-bis(acetatomercurimethyl)dioxane (BAMD). Fractions were investigated in their analytical CsCl profiles and a number of specific sequences were localized in them. The results so obtained led to an improved understanding of the organization of nucleotide sequences in the human genome, as well as to the discovery that a class of DNA having a very high G + C content and not represented in the mouse genome, is particularly rich in genes and interspersed repetitive sequences.

Base Sequence↗
PubMed1986

Isolation of intercalator-dependent protein-linked DNA strand cleavage activity from cell nuclei and identification as topoisomerase II.

DNA intercalating agents such as 4'-(9-acridinylamino)methanesulfon-m-anisidide (m-AMSA) have previously been found to induce in mammalian cells the formation of protein-associated DNA single- and double-strand breaks. In the current work, an activity characterized by the production of DNA-protein links associated with DNA strand breaks and by stimulation by m-AMSA was isolated from L1210 cell nuclei and was shown to be due to topoisomerase II. Nuclei were extracted with 0.35 M NaCl, and the extract was fractionated by gel filtration, DNA-cellulose chromatography, and glycerol gradient centrifugation. A rapid filter binding assay was devised to monitor the fractionation procedure on the basis of DNA-protein linking activity. The active DNA-cellulose fraction contained both topoisomerase I and topoisomerase II whereas the glycerol gradient purified material contained only topoisomerase II activity. The properties of the active material were studied at both stages of purification. m-AMSA enhanced the formation of complexes between purified topoisomerase II and SV40 DNA in which the DNA sustained a single- or double-strand cut and the enzyme was covalently linked to the 5' terminus of the DNA. This action was further enhanced by ATP, as well as by nonhydrolyzable ATP analogues. m-AMSA inhibited the topoisomerization and catenation reactions of topoisomerase II, probably because of trapping of the enzyme-DNA complexes. The activity showed a dependence on the type of DNA intercalators used, analogous to what was previously observed in intact cells. m-AMSA had no effect on topoisomerase I.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1985

The mosaic genome of warm-blooded vertebrates.

Most of the nuclear genome of warm-blooded vertebrates is a mosaic of very long (much greater than 200 kilobases) DNA segments, the isochores; these isochores are fairly homogeneous in base composition and belong to a small number of major classes distinguished by differences in guanine-cytosine (GC) content. The families of DNA molecules derived from such classes can be separated and used to study the genome distribution of any sequence which can be probed. This approach has revealed (i) that the distribution of genes, integrated viral sequences, and interspersed repeats is highly nonuniform in the genome, and (ii) that the base composition and ratio of CpG to GpC in both coding and noncoding sequences, as well as codon usage, mainly depend on the GC content of the isochores harboring the sequences. The compositional compartmentalization of the genome of warm-blooded vertebrates is discussed with respect to its evolutionary origin, its causes, and its effects on chromosome structure and function.

Animals↗
PubMed1983

Reconstitution of intercalator-induced DNA scission by an active component from nuclear extracts.

Treatment with intercalating agents causes formation of protein-associated DNA breaks in mammalian cells in culture and in the nuclei isolated from these cells. We found that this effect, when induced by the intercalator m-AMSA, required a component which could be dissociated from nuclei by 0.3 M NaCl. The effect was restored by combining the extracted nuclei with the nuclear extract. The active component of the extract eluted in gel filtration at a point corresponding to a molecular weight of 800 000. During its reaction with DNA, DNA-protein links and DNA breaks appeared in approximately equal frequencies. In this respect the reaction stimulated by m-AMSA resembled the reaction of a topoisomerase with DNA. However, intercalator-stimulated formation of protein-associated DNA breaks differed from the activity of the nuclear topoisomerase I in that there was a different optimum salt concentration and a different apparent molecular weight.

Aminoacridines↗
PubMed1983

Competitive inhibition of nicking--closing enzymes may explain some biological effects of DNA intercalators.

Intercalating agents cause varied and multiple biological effects. These include the inhibition of RNA and DNA synthesis, frameshift mutations and protein-associated DNA breaks. However, some non-intercalating analogs of intercalating compounds behave similarly. The model of DNA intercalation does not adequately explain all these biological effects. It is suggested here that intercalators and similar compounds may competitively inhibit the closing reaction of some nicking--closing enzymes. Hypothetical mechanisms built on this suggestion are presented for the formation of protein associated DNA breaks, frameshift mutation, inhibition of macromolecular synthesis, and recombination.

Animals↗
PubMed1983

Differential crosslinking of histones and non-histones in nuclei by cis-Pt(II).

When nuclei were treated with the chemotherapeutic agent, cis-Pt(II), they were crosslinked to the extent that their nuclear morphology as assayed by light microscopy was retained even in the presence of SDS. Protein analysis showed that the histones were completely absent from these nuclear structures, while the non-histone proteins, with one possible exception, were completely retained. When the nuclear structures in SDS were treated with thiourea to reverse the crosslinks, the non-histone proteins were liberated and the nuclear structures disappeared. When treated with Proteinase K in SDS, the nuclear structures also disappeared, indicating that protein components were necessary to maintain the structures.

Animals↗
PubMed1983

Topoisomerase I activities in L-1210 cell nuclei.

Topoisomerase was extracted from L-1210 nuclei by 0.35 M sodium chloride Extract fractionation by molecular weight revealed two activities. The activities were quantitatively determined using a technique based on the electrophoretic separation of 3H-labelled DNA product from substrate. This technique enables the determination of topoisomerase activity in extracts where nuclease activity is also present.

Animals↗
PubMed1982

Ellipticine-induced protein-associated DNA breaks in isolated L1210 nuclei.

DNA intercalating agents have been found to produce protein-associated DNA strand breaks in mammalian cells. At a first step towards a subcellular system for the study of this reaction, we demonstrate that the reaction can take place in isolated cell nuclei. Ellipticine induces in these nuclei DNA strand breaks and stable DNA-protein complexes. Complexes and breaks are present in equivalent amounts. DNA breaks are revealed only if protein-mediated DNA adsorption to filters is abolished. These findings make it unlikely that similar effects observed in cells in culture after treatment with intercalating agents are caused by metabolically activated drugs.

Alkaloids↗
PubMed1982

Metrizamide gradient centrifugation of histone-DNA complexes.

Mouse DNA was complexed with different combinations of histones. Most of these complexes were separated in metrizamide gradient into two fractions--heavy and light. At the protein: DNA ratio of 0.8 a complex of DNA with five histones contained 60% of DNA in the heavy fraction whereas in the absence of histone H1, this fraction contained only 40% of DNA. Complexes of DNA with the H2A + H2B histone pair banded in metrizamide as one, light fraction at the density of 1.140 g/cm3. DNA reconstituted with the H3 + H4 histone pair formed in metrizamide two peaks of 1.240 g/cm3 and 1.140 g/cm3. Among complexes of DNA with single histones H2A, H2B, H3 and H4 only the complex DNA-H4 formed a component having in metrizamide a density of 1.200 g/cm3, similar to the density of purified chromatin.

Animals↗
PubMed1980

The nature of inactivating lesions produced by platinum(II) complexes in phage lambda DNA.

Lambda DNA loses transfectivity and acquires interstrand cross-links after treatment with either trans-Pt(II) or cis-Pt(II). With trans-Pt(II) there is close to an equivalence between the fraction of lambda DNA cross-linked and the fraction inactivated. In contrast, with cis-Pt(II) there are approx. 5 inactivating lesions for each lambda DNA interstrand cross-link. These results suggested that trans-PT(II) does not introduce intrastrand inactivating lesions into lambda DNA while cis-Pt(II) does so. To verify this conclusion, the cross-linked and uncross-linked fractions of lambda DNA treated with trans-PT(II) or cis-Pt(II) were separated on alkaline sucrose gradients. After trans-Pt(II) treatment, the uncross-linked fraction of lambda DNA was transfective when renaturated. However after cis-Pt(II) treatment the uncross-linked fraction of lambda DNA was not transfective when renatured. Thiourea treatment restored transfectivity to all inactivated fractions, showing that these lesions are reversible. We conclude that trans-Pt(II) inactivates lambda DNA primarily by introducing interstrand cross-links but that cis-Pt(II), although it also introduces interstrand cross-links, inactivates lambda DNA primarily by introducing intrastrand lesions.

Bacteriophage lambda↗
PubMed1979

Thiourea reverses cross-links and restores biological activity in DNA treated with dichlorodiaminoplatinum (II).

Cis and trans dichlorodiaminoplatinum (II) compounds bind to DNA and form DNA cross-links, which are usually considered to be irreversible. Thiourea can reverse these cross-links without any apparent breakdown of the DNA. In addition, cis- and trans-Pt (II) treatment of lambda decreases its transfectivity. After suitable incubation with thiourea, full transfectivity of Pt(II)-treated lambda DNA can be restored.

Animals↗
PubMed1978

Complex of DNA with chromatin proteins investigated by isopycnic centrifugation in metrizamide.

Complexes of mouse main band DNA with a fraction of non-histone proteins (NHP), having a high affinity for DNA, in the absence or presence of histones have been investigated by gradient centrifugation in metrizamide. Two types of complexes were formed at an input ratio of NHP to DNA between 1 and 2.5. In metrizamide gradients a majority of DNA was found in the light complex (at the density of 1.14-1.16 g/cm3) even at the very high NHP to DNA ratio. When histones were present in the reaction mixture, most of the DNA was found in the heavy complex (1.19-1.21 g/cm3). The electrophoretic profiles of the proteins recovered from the heavy and light complexes were different; some fractions of nonhistone proteins were present only in the heavy component.

Animals↗