We previously reported the identification of a novel nuclear compartment detectable in heatshocked HeLa cells that we termed stress-induced Src-activated during mitosis nuclear body (SNB). This structure is the recruitment center for heat shock factor 1 and for a number of RNA processing factors, among a subset of Serine-Arginine splicing factors. In this article, we show that stress-induced SNBs are detectable in human but not in hamster cells. By means of hamsterϾhuman cell hybrids, we have identified three human chromosomes (9, 12, and 15) that are individually able to direct the formation of stress bodies in hamster cells. Similarly to stress-induced SNB, these bodies are sites of accumulation of hnRNP A1-interacting protein and heat shock factor 1, are usually associated to nucleoli, and consist of clusters of perichromatin granules. We show that the p13-q13 region of human chromosome 9 is sufficient to direct the formation of stress bodies in hamsterϾhuman cell hybrids. Fluorescence in situ hybridization experiments demonstrate that the pericentromeric heterochromatic q12 band of chromosome 9 and the centromeric regions of chromosomes 12 and 15 colocalize with stress-induced SNBs in human cells. Our data indicate that human chromosomes 9, 12, and 15 contain the nucleation sites of stress bodies in heat-shocked HeLa cells.

Griseofulvin (GF) is a mycotoxin produced by various species of Penicillium including P. griseofulvum Dierckx, P. janczewski (P. nigricans) and P. patulum. It is active against dermatophytic fungi of different species in the genera Microsporum, Trychophyton and Epidermophyton. Because of its capacity to concentrate in the keratinous layer of the epidermis and its relatively low toxicity in man, it has been extensively used in the therapy of dermatophytoses by oral administration. The biological activity of GF towards fungi is manifested as nuclear and mitotic abnormalities followed by distortions in the hyphal morphology. Mitotic segregation is also induced in fungi by GF treatment. In higher eukaryotes the cytostatic action of GF is essentially due to a mitotic arrest at late metaphase/early anaphase. The cytological effects observable both in vivo and in vitro on different plant and animal cell systems, include C-mitoses, multipolar mitoses and multinuclearity. Prolonged GF treatment in experimental animals provokes biochemical changes consisting mainly of disturbances of porphyrin metabolism, variation in the microsomal cytochrome levels and formation of Mallory bodies. In mice these alterations are followed by the development of multiple hepatomas. Evidence of tumor induction by GF has been obtained in mice and rats, but not in hamsters. GF may also act either as a promoting or a co-carcinogenic agent, depending on the circumstances of its administration. It has been found to increase the frequency of cell transformation induced by polyoma virus, but not to induce cell transformation per se. Induction of sperm abnormalities has been observed in GF-treated mice. The embryotoxic and teratogenic action of GF has been demonstrated in pregnant rats exposed during organogenesis. Genetic effects of GF have been investigated by the following tests: Salmonella/microsome mutagenicity assay, point mutations in mammalian and plant cells, DNA damage and repair, SCE, chromosome aberrations, micronuclei, dominant lethals, aneuploidy in lower and higher eukaryotes. A positive response has been obtained in the assays on numerical chromosome changes in all the systems analyzed; limited or inconclusive evidence has been obtained for SCE and structural chromosome changes. Doubled or highly polyploid sets can be detected in all types of cells during or immediately after GF treatment. A marked increase in chromosome number variation is observed at various times after withdrawal of the drug, with prevailing hyperdiploid and reduced sets in animal cells and plant cells respectively.(ABSTRACT TRUNCATED AT 400 WORDS)

Five human glioblastoma cell lines were analyzed for oncogene activation with a panel of probes. Abnormal expression of the epidermal growth factor receptor (EGFr) gene was detected in four of five lines; N-ras oncogene overexpression was found in all five cell lines. These results were subsequently confirmed with fresh brain tumor and nonneoplastic brain tissue biopsy samples; increased expression of the N-ras proto-oncogene was observed in five of five glioblastomas, all of which also showed EGFr gene overexpression, but not in well-differentiated gliomas or in nonneoplastic brain tissue specimens. No significant differences in Ha-ras and Ki-ras expression were observed. Preliminary histochemical observations showed that intracellular levels of transforming growth factor alpha, a putative biochemical link between these two oncogenes, were significantly higher in glioblastoma cells than in controls.

A human supernumerary minichromosome (MC), previously identified as a derivative of chromosome 9, has been introduced into Chinese hamster ovary (CHO) cells by means of cell fusion. A hybrid clone containing the MC as the only free human chromosome was isolated. A selectable marker gene (neo) inserted into a yeast artificial chromosome (YAC) has been successfully targeted to the MC centromeric DNA via co-transfection with chromosome-9-specific alpha satellite DNA. In situ hybridization and Southern blotting experiments demonstrated that the intact neo gene was integrated into the MC centromeric DNA. Studies on the clonal distribution and on the stability of the MC either in the presence or in the absence of the selective agent have been carried out. The MC is susceptible to further manipulations and may thus represent a model for the construction of a large-capacity vector for somatic gene therapy.

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