The isolation and characterization of a clonally related series of murine retrovirus-infected mouse cells. (65/106)

Starting with cloned NIH 3T3 mouse cells we have isolated a series of related lines infected with the Kirsten murine sarcoma/leukaemia (MSV/MLV) virus complex. These lines exhibit all three possible infected cell phenotopes: (i) transformed MSV/MLV producers; (ii) non-transformed MLV producers; (iii) transformed non-producers. We have also selected non-transformed revertants from one of the non-producer clones. This series of lines allows the study of the expression of the virus genome against a constant background of cellular gene expression. We have further characterized the lines with regard to anchorage dependence of growth, tumorigenicity and the presence of a rescuable sarcoma genome. The non-producer clones are uniform in their transformed properties. The revertants contain rescuable sarcoma virus, biologically indistinguishable from the original transforming virus, implying that the reversion is due to a change in cellular rather than viurs genetic information.  (+info)

Virus RNA species in kirsten murine sarcoma virus-transformed mink cells. (66/106)

The nuclear and cytoplasmic RNAs from Kirsten murine sarcoma virus (KiMuSV)-transformed non-producer mink cells were studied for the species of virus-specific RNA by fractionation in agarose gels, transfer to diazotized aminophenylthioether paper and hybridization to complementary DNA probe. In both nuclei and cytoplasm, only genome-length KiMuSV-specific RNA was detected. No subgenomic virus RNA species was detected in poly(A+) or poly(A-) RNA fractions. The same observations were made in KiMuSV-transformed mink cells superinfected with feline leukaemia viruses. The significance of these findings is discussed.  (+info)

Viral susceptibility of skin fibroblasts from patients with Huntington disease. (67/106)

Cultured skin fibroblasts from patients with Huntington disease (HD) and age-matched controls were tested for susceptibility to vesicular stomatitis virus (VSV) and transformation by Kirsten mouse sarcoma virus (KiMSV). The HD and control cells could not be distinguished on the basis of viral replication, plaque morphology, virus yield, or susceptibility to transformation by KiMSV. These findings suggest that the HD gene product, if expressed within peripheral tissue, does not selectively alter or interfere with viral replication.  (+info)

Morphological and functional differentiation of Kirsten murine sarcoma virus-transformed rat adrenocortical cell lines. (68/106)

This study was undertaken to examine relationships of the phenotype of malignant cells to target cell properties and to events early in the transformation process. Eighteen transformed lines were obtained by Kirsten murine sarcoma virus infection of cells from adrenal glands of rats ages 4 to 30 weeks, at first or second passages in culture. They were grown either as fibroblastic adrenocortical stem cells or as more differentiated epithelial cells, depending on culture conditions. Of 14 lines examined for their capacity to synthesize corticosteroids, 11 converted [14C]pregnenolone to progesterone, and one converted to deoxycorticosterone. In vivo, seven lines produced tumors resembling pleomorphic carcinomas, six lines grew as sarcomas, four grew as mixed tumors, and one line produced anaplastic tumors. Distinguishing features in culture of the carcinoma-producing lines were early onset and rapid progression of morphological transformation, a noncohesive epithelial cell form in some lines, lack of extracellular matrix, and, possibly, and origin in older animals. In contrast, sarcoma-producing cells were fibroblastic and cohesive, produced extracellular matrix, and transformed morphologically after longer and less well-defined periods in culture. The variation in histopathology was unrelated to the differentiation of the target cells and to the capacity of the transformed cells to synthesize corticosteroids. The results show that adrenocortical cells, transformed by Kirsten murine sarcoma virus after short-term culture, usually retain some functional differentiation and sometimes resemble human adrenocortical carcinomas histologically. The susceptibility of adrenocortical cells to Kirsten murine sarcoma virus raises the possibility that mesodermally derived epithelia in general may be target tissues for C-type sarcoma viruses.  (+info)

Transformation of cultured rat ovarian surface epithelial cells by Kirsten murine sarcoma virus. (69/106)

A method is described for the culture of rat ovarian surface epithelial cells, i.e., the cellular component thought to be the source of most ovarian cancers. These cells in culture have a characteristic epithelial morphology which distinguishes them from other ovarian cell types. Cultured surface epithelial cells are histochemically positive for 17 beta-hydroxysteroid dehydrogenase and negative for delta 5-3 beta-hydroxysteroid dehydrogenase, the same as in cryostat sections of whole rat ovary. Ultrastructurally, cultured surface epithelial cells have basement membranes, microvilli, and apical intercellular junctions. Kirsten murine sarcoma virus was used to produce three transformed cell lines from pure first-passage cultures of these cells. These three lines retained 17 beta-hydroxysteroid dehydrogenase activity and showed slight delta-3 beta-hydroxysteroid dehydrogenase activity. Tumors resulting when these cells were injected s.c. or i.p. into immunosuppressed female rats were highly malignant, resembling histologically human ovarian endometrioid stromal sarcoma. This is the first demonstration of the susceptibility of ovarian surface epithelium to an oncogenic virus.  (+info)

Structure and functions of the Kirsten murine sarcoma virus genome: molecular cloning of biologically active Kirsten murine sarcoma virus DNA. (70/106)

The unintegrated closed circular form of viral DNA prepared from NIH3T3 cells infected with Kirsten murine sarcoma virus was cloned into bacterial plasmid pBR322. The closed circular DNA, which consisted of two different-sized populations, was enriched from the virus-infected cells, linearized with BamHI, and inserted into pBR322 DNA. Four different recombinant DNAs (clones 2, 4, 6, and 7) were obtained, and a physical map of each was constructed by using various restriction enzymes. Clone 4 DNA had the largest insertion, corresponding to a complete copy of the linear DNA. This suggested that this insertion contained two copies of the 0.55-kilobase pair long terminal redundant sequence. Clone 2 and clone 6 insertion DNAs had deletions of 0.2 and 0.5 kilobase pair, respectively, which mapped near the right end (3' side of viral RNA) of the linear DNA. Clone 7 DNA appeared to have a deletion of a single copy of the large terminal redundant sequence. Transfection of BALB3T3 cells with the clone 4 DNA insertion showed that this DNA had transforming activity. The efficiency of transfection with clone 4 Kirsten murine sarcoma virus DNA was enhanced eightfold by inserting EcoRI-cleaved viral DNA into the EcoRI site of pBR322. The EcoRI-inserted DNA produced foci with single-hit kinetics, suggesting that a single molecule of Kirsten murine sarcoma virus DNA can induce transformation. Results of transfections with EcoRI-inserted Kirsten murine sarcoma virus DNA cleaved with various restriction enzymes suggested that the first 3.3-kilobase pair region at the left end of the linear DNA is important for the initiation of transformation or maintenance of transformation or both.  (+info)

Identification of unintegrated forms of Kirsten murine sarcoma viral DNA and restriction endonuclease cleavage map of linear DNA. (71/106)

We detected unintegrated linear 7.0-kilobase pair DNA and covalently closed circular DNA species in NIH3T3 cells recently infected with Kirsten murine sarcoma virus. Using the linear DNA, we constructed a restriction endonuclease cleavage map and compared it with the map of Harvey murine sarcoma virus. The restriction endonuclease maps of two segments, one 1.2 kilobase pairs (SmaI site) to 3.7 kilobase pairs (HindIII site) from the right end (corresponding to the viral 3' side) and the other 0.5 kilobase pair (SmaI and KpnI sites) to 0.9 kilobase pair (KpnI site) from the left end, were identical in the two virus types.  (+info)

Analysis of the myeloproliferative sarcoma virus genome: limited changes in the prototype lead to altered target cell specificity. (72/106)

The myeloproliferative sarcoma virus (MPSV) derived from Moloney sarcoma virus (MSV-Mol) is a unique sarcoma virus which causes expansion of the hematopoietic stem cell compartment as well as the erythroid and myeloid cell lineages. MPSV also induces spleen focus formation in adult mice as do Friend and Rauscher viruses. Analysis of the MPSV genome on methyl mercury gels showed that the genome size is 7.0 kilobases, which is larger than the defective genome of any known MSV-Mol isolate. Hybridization analysis with specific cDNA probes showed that MPSV is a modified sarcoma virus with no sequences in the unique region of the defective sarcoma genome related to unique Friend virus sequences. The only viral sequences in the defective genome other than helper virus-related sequences are derived from the Moloney sarcoma virus genome with no new cellular sequences added. There was no evidence for induction of xenotropic virus sequences in MPSV-infected spleens of DBA/2J mice, indicating that spleen focus formation can be obtained by different mechanisms.  (+info)