Evidence for a new tumour suppressor locus (DBM) in human B-cell neoplasia telomeric to the retinoblastoma gene.

Roughly 25% of human B-cell chronic lymphocytic leukaemias (CLL) are characterized by a chromosomal lesion involving 13q14. This region contains the retinoblastoma gene (RB1). We have used a variety of techniques to determine whether RB1 or some other locus is the critical region in 11 cases of low grade B-cell malignancy (mainly CLL), all with deletions or translocations involving 13q14. In all cases, except the one with minimal disease, there was deletion or a structural lesion in the region of D13S25, with at least 4 cases showing homozygous disruption. We conclude that D13S25 lies close to a tumour suppressor locus whose inactivation contributes to the initiation or progression of low grade B-cell malignancy. This locus is located at least 530 kilobases telomeric to RB1.

A familial RB1 mutation detected by the HOT technique is homozygous in a second primary neoplasm.

Using the hydroxylamine-osmium tetroxide (HOT) technique, we have identified a constitutional point mutation in the retinoblastoma susceptibility gene (RB1) which segregates with the expression of retinoblastoma in five affected family members. One member developed a second primary tumor, a small-cell lung carcinoma (SCLC), which metastasized to the liver. Analysis of liver tumour DNA revealed homozygosity for the constitutional mutation, a G----A transition at the fifth base of intron 21, resulting in the excision of exon 21 from the mRNA. This is the first demonstration of homozygotization of a constitutional RB mutation in a metastatic second primary tumour and underlines the usefulness of the HOT technique for identification of mutations of the RB1 gene.

Karyotypic abnormalities and immunoglobulin gene rearrangements in Hodgkin's disease.

Biopsy samples from seven patients with Hodgkin's disease (HD) were examined for cytogenetic abnormalities and rearrangement of the genes encoding the immunoglobulin chains and T-cell receptor chains. Three samples demonstrated clonal rearrangements of both IgH and IgL genes. No rearrangements of the TCR beta genes were detected in any of the samples. Karyotypic abnormalities were also found but only in the three cases where a clonal rearrangement of the immunoglobulin genes was shown. Two of these three cases had multiple karyotypic abnormalities, with the remaining patient being trisomic for chromosome 16 as the sole abnormality. These results are discussed and compared with previous reports in the literature concerning HD.

HRAS1-selected, chromosome-mediated transformants vary in phenotype in vitro and tumorigenic potential in vivo.

Transfection of mouse C127 cells with mitotic chromosomes isolated from a human EJ bladder carcinoma cell line gave rise, at high frequency, to foci of transformed cells. Independent, HRAS1-selected chromosome-mediated transformants displayed distinctive cellular morphologies in monolayer culture and colony-forming abilities in low-melting-point agarose. Subcutaneous inoculation of neonatally thymectomized, Ara-C-protected, total-body-irradiated CBA mice was used to compare the tumorigenic potential of each transformant. Significant quantitative and qualitative differences in tumorigenicity were found between transformants which correlated with differences in malignant phenotype observed in vitro. The sensitivity of the tumorigenicity assay is such that rare transformation events can be selected directly in vivo.

Developmental changes in translatable RNA species associated with meiosis and spore formation in Saccharomyces cerevisiae.

During Saccharomyces cerevisiae sporulation distinct changes in translatable mRNA species have been detected by two-dimensional gel electrophoresis of the polypeptides produced in a messenger-dependent, cell-free rabbit reticulocyte lysate primed with RNA prepared from a/alpha, a/a, and alpha/alpha isogenic diploids at different stages of sporulation. The availability of functional mRNA increased by about 25% during the first 4 h after transfer of either sporulating (a/alpha), or nonsporulating (a/a and alpha/alpha) diploids to sporulation medium. Thereafter functional mRNA decreased such that in the a/alpha strain after 24 h there was only about 50% of the amount in vegetative cells; a less marked decrease was observed in the a/a and alpha/alpha strains. Of 750 mRNA species detected, 43 underwent alterations only during sporulation in the a/alpha strain, whereas 36 changes were common to all three strains and one mRNA specific to alpha/alpha vegetative cells was detected. Only four of the sporulation-specific changes were due to the de novo appearance of translatable species, and two of these became predominant species of the total population. The majority of the specific changes were due to either permanent or transient increases in the concentration of individual mRNA species; 11 decreases were found. Changes were found at most stages of sporulation, although many occurred in either of two stages, one early (before 2 h) and the other later (between 6 and 8 h) when commitment to meiotic segregation was beginning. The results provide evidence for both quantitative and, to a lesser extent, qualitative transcriptional control of gene expression during sporulation.