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R Sheinin

Publications and source records attributed to R Sheinin.

13 recordsLinked to original sources
PubMed1978

Some properties of chromatin synthesized by mouse-L-cells temperature-sensitive in DNA replication.

When ts A1S9 mouse L-cells are incubated at the nonpermissive temperature (38.5 degrees) DNA synthesis proceeds at the normal rate for 6 to 8 h; it then declines to attain 1 to 5% of this rate after 24 h. General protein synthesis from precursor leucine is relatively unaffected by the high temperature. In contrast, protein formation from lysine (and arginine) remains unchanged for 12 to 15 h after temperature upshift. It then drops and plateaus at about 25% of the initial rate after 32 h. The chromatin protein and DNA are fully conserved in ts A1S9 cells incubated at 38.5 degrees for at least 24 h after full expression of the ts defect. Temperature inactivation of the ts A1S9 gene product results in inhibition of de novo formation of chromatin. This is evidenced by coordinate suppression of incorporation of dThd and of lysine and arginine into chromatin-bound DNA and histone, respectively.

Chromatin↗
PubMed1978

Structure of interphase nuclei in relation to the cell cycle. Chromatin organization in mouse L cells temperature-sensitive for DNA replication.

Mutant lines of mouse L cells, TS A1S9, and TS C1, show temperature-sensitive (TS) DNA synthesis and cell division when shifted from 34 degrees to 38.5 degrees C. With TS A1S9 the decline in DNA synthesis begins after 6-8 h at 38.5 degrees C and is most marked at about 24 h. Most cells in S, G2, or M at temperature upshift complete one mitosis and accumulate in the subsequent interphase at G1 or early S as a result of expression of a primary defect, failure of elongation of newly made small DNA fragments. Heat inactivation of TS C1 cells is more rapid; they fail to complete the interphase in progress at temperature upshift and accumulate at late S or G2. Inhibition of both cell types is reversible on return to 34 degrees C. Cell and nuclear growth continues during inhibition of replication. Expression of both TS mutations leads to a marked change in gross organization of chromatin as revealed by electron microscopy. Nuclei of wild-type cells at 34 degrees and 38.5 degrees C and mutant cells at 34 degrees C show a range of aggregation of condensed chromatin from small dispersed bodies to large discrete clumps, with the majority in an intermediate state. In TS cells at 38.5 degrees C, condensed chromatin bodies in the central nuclear region become disaggregated into small clumps dispersed through the nucleus. Morphometric estimation of volume of condensed chromatin indicates that this process is not due to complete decondensation of chromatin fibrils, but rather involves dispersal of large condensed chromatin bodies into finer aggregates and loosening of fibrils within the aggregates. The dispersed condition is reversed in nuclei which resume DNA synthesis when TS cells are downshifted from 38.5 degrees to 34 degrees C. The morphological observations are consistent with the hypothesis that condensed chromatin normally undergoes an ordered cycle of transient, localized disaggregation and reaggregation associated with replication. In temperature-inactivated mutants, normal progressive disaggregation presumably occurs, but subsequent lack of chromatin replication prevents reaggregation.

Cell Count↗
PubMed1978

Mutant mouse L-cells: a model for megaloblastic anaemia.

When temperature-sensitive (ts) mutant lines of mouse L-cells, ts AIS9 and ts CI, are shifted from 34C to 38.5C, a rapid inhibition of DNA synthesis and mitosis occurs. During this phase, cell and nuclear growth continues and results in a substantial increase in cell and nuclear volume. Such cellular modifications are also associated with a marked dispersal of the condensed chromatin masses of interphase nuclei, so that after 48-72 h of incubation at 38.5C, nuclear profiles of both ts cell lines bear a striking resemblance to the nuclear features characteristic of megaloblastic anaemia. Despite these marked alterations in nuclear chromatin organization, morphometric analysis indicates that the volume of condensed chromatin does not decrease. Current biochemical, cytological and morphometric data on the two ts lines of mutant mouse L-cells during expression of the mutation, suggest that they might provide a useful model to further elucidate cytological features of megaloblastic anaemia.

Anemia, Macrocytic↗
PubMed1977

Semi-conservative and non-conservative replication of DNA in temperature-sensitive mouse L-cells.

The mode of DNA replication has been studied in wild-type mouse L-cells (WT-4) and in two subclones (TS A1S9 and ts C1 cells) which are temperature-sensitive in DNA synthesis. It has been demonstrated that DNA is replicated by the semi-conservative mechanism in WT-4 cells grown at 34 degrees C or at 38.5 degrees C throughout the logarithmic phase and into the stationary phase. Similar results were obtained with ts A1S9 and ts C1 cells grown at the permissive temperature (34 degrees C). When the latter cells were incubated at the non-permissive temperature (38.5 degrees C) inactivation of DNA synthesis appeared to proceed through three general stages. During the first 24 h after temperature upshift suppression of semi-conservative DNA replication occurred. During the second stage a very low level of semi-conservative synthesis was maintained. During the third stage, incorporation of dThd into DNA began to increase, often reaching 10-20% of control levels after 3-5 days. During this third stage DNA synthesis was effected by a non-conservative mechanism. Temperature-inactivated ts A1S9 cells and ts C1 cells were able to perform semi-conservative synthesis upon back-shift to 34 degrees C, using as template that DNA synthesized prior to temperature upshift.

Centrifugation, Density Gradient↗
PubMed1977

Mitochondrial DNA synthesis in mouse L cells temperature sensitive in nuclear DNA replication.

Temperature-sensitive (ts) A 1S9 mouse L cells continue to synthesize double-stranded covalently closed mitochondrial (mt) DNA at a temperature (38.5 degrees C) which is nonpermissive for chromosomal DNA replication. The amount of mt DNA made appears to be quantitatively linked to nuclear DNA synthesis. Nuclear DNA replication proceeds normally for 6-8 h after the cells are shifted to 38.5 degrees C, and then declines to reach a minimum at 20-24 h. The level of mt DNA synthesis remains high during this period and decreases once the ts lesion has been established.

Animals↗
PubMed1976

Biosynthesis of plasma membrane components by SV40-virus-transformed 3T3 mouse cells temperature sensitive for expression of some transformed cell properties.

We have studied the plasma membranes of an SV40-transformed 3T3 cell line temperature sensitive for the transformed growth phenotype (ts H6-15 cells), and have found that they vary little as a function of temperature of cultivation. Analysis by polyacrylamide gel electrophoresis was performed on plasma membranes prepared from ts H6-15 cells cultured at the permissive (32 degrees C) and non-permissive (39 degrees C) temperatures and radioactively-labelled in several ways. No significant differences were seen when the electrophoretic patterns of polypeptides of the plasma membranes of ts H6-15 cells, grown through 3-4 generations in medium containing radioactive leucine (32 degrees C and 39 degrees C temperatures) were compared. Plasma membranes derived from cells similarly grown in medium with radioactive glucosamine indicated that extensive alterations in the intrinsic glycopeptides occurred in association with alteration in growth phenotype. A shift towards decreased synthesis of large molecular weight (congruent to 100 000-160 000) glycopeptides occurred in cells grown at the temperature of non-transformed growth (39 degrees C). A decrease in amount of a 120 000 molecular weight glycopeptide at 39 degrees C was the most prominent of these alterations. We have studied the surface exposure of polypeptides and glycopeptides of intact cells grown at 32 and 39 degrees C, using lactoperoxidase-catalyzed iodination, NaBH4 reduction of galactose oxidase-treated cells, and metabolic-labelling with glucosamine of trypsin-sensitive molecules. We found no major qualitative differences between whole cell extracts or between plasma membrane preparations of cells cultivated at the permissive and non-permissive temperatures. Of special interest was the observation that the formation and surface exposure of a trypsin-sensitive, 240 000 molecular weight polypeptide appeared not to be ts in ts H6-15 cells. The significance of these observations will be discussed.

Cell Fractionation↗
PubMed1976

A consideration of the role of cell surface macromolecules in the process of viral transformation.

There is extensive physiological evidence implicating the cell surface as the key organelle which mediates the cell:cell interactions which underlie both normal and neoplastic growth. This information has now been supplemented with biochemical and biophysical data which indicates that surface macromolecules, in particular the heteroglycans of transformed cells, differ from those which lie at the periphery of normal cells. In the case of cells neoplastically transformed by most tumour viruses it is clear that the small virus genome (2-5 x 10(6) daltons) cannot carry the total genetic information to accomodate these various biochemical modifications, if indeed they are encoded in separate genes (1). To examine the part played in transformation by cellular genes coding for surface heteroglycan formation, we have turned to a study of SV-3T3 cells (ts H6-15) which are temperature-sensitive for expression of the transformed cell phenotype (2). The data show that cells grown under conditions permissive and non-permissive for such expression exhibit the same pattern of formation of glycolipids, and the majority of the polypeptides of the plasma membrane. There are, however, significant differences in the synthesis of some glycopeptides. A large molecular weight, trypsin-labile glycopeptide, present at the surface of untransformed fibroblasts but barely measurable in some of their virus-transformed derivatives (3), was detected, essentially at the same level, at the surface of ts H6-15 cells grown at the permissive and non-permissive temperatures. The signficance of these observations is discussed.

Cell Adhesion↗
PubMed1976

Preliminary characterization of the temperature-sensitive defect in DNA replication in a mutant mouse L cell.

Temperature-sensitive ts A1S9 mouse L cells synthesize DNA apparently normally for 6-8 hr upon incubation at 38.5 degrees C. Thereafter, these cells are able to perform limited polydeoxyribonucleotide chain synthesis at the high temperature, but are unable to convert newly replicated small single-strand segments of DNA (of the order of molecular weight 10(6) daltons) to large molecular weight chromosomal DNA. Data obtained are compatible with a model which suggests that ts A1S9 cells are able to carry out most individual reactions of DNA synthesis at the high temperature, but are temperature-sensitive in a protein which participates in the joining of small DNA segments to make chromosomal DNA strands. When cells are reincubated at a permissive temperature, after the temperature-sensitive lesion has been established, they recover the latter capability several hours before they are able once again to synthesize DNA at normal rates.

Centrifugation, Density Gradient↗
PubMed1975

Viruses: causative agents of cancer.

Evidence accumulated over a period of 60 years has clearly established that a number of different viruses cause neoplasia (of a broad spectrum) amongst a variety of animals. Although it is known that viruses do produce self-limiting proliferative human diseases, complete verification of a human viral carcinogen remains to be provided. This paper presents an overview of the conceptual and practical tools for the detection of tumor viruses, which have derived from experimental model systems. It indicates how these are being applied to studies of human malignant disease, with special reference to assessing a possible viral etiology for laryngeal cancer.

Adenoviridae↗