Identification of cadmium-induced mutations in a mammalian cell line

Epidemiological studies have shown cadmium to induce cancer in humans, while experimental studies have proven this metal to be a potent tumor inducer in animals. However, cadmium appears nonmutagenic in most prokaryotic and eukaryotic mutagenesis assays. In this study, we present the identification of mutations in normal rat kidney cells infected with the mutant MuSVts110 retrovirus (6m2 cells) as a result of treatment with cadmium chloride. The detection of these mutations was facilitated by the use of a novel mutagenesis assay established in this laboratory. The 6m2 reversion assay is a positive selection system based on the conditional expression of the MuSVts110 v-mos gene. In MuSVts110 the gag and mos genes are fused out of frame, thus the translation of the v-mos sequence requires a frameshift in the genomic RNA. In 6m2 cells this frameshift is accomplished by the temperature-dependent splicing of the primary MuSVts110 transcript. Splicing of MuSVts110, which is mediated by cis-acting sequences, occurs when 6m2 cells are grown at 33$\sp\circ$C and below, but not at 39$\sp\circ$C. Therefore, 6m2 cells appear transformed at low growth temperatures, but take on a morphologically normal appearance when grown at high temperatures. The treatment of 6m2 cells with cadmium chloride resulted in the outgrowth of a number of cells that reverted to the transformed state at high growth temperatures. Analysis of the viral proteins expressed in these cadmium-induced 6m2 revertants suggested that they contained mutations in their MuSVts110 DNA. Sequencing of the viral DNA from three revertants that constitutively expressed the P85$\sp{gag{-}mos}$ transforming protein revealed five different mutations. The Cd-B2 revertant contained three of those mutations: an A-to-G transition 48 bases downstream of the MuSVts110 3$\sp\prime$ splice site, plus a G-to-T and an A-to-T transversion 84 and 100 bases downstream of the 5$\sp\prime$ splice site, respectively. The Cd-15-5 revertant also contained a point mutation, a T-to-C transition 46 bases downstream of the 5$\sp\prime$ splice site, while Cd-10-5 contained a three base deletion of MuSVts110 11 bases upstream of the 3$\sp\prime$ splice site. A fourth revertant, Cd-10, expressed a P100$\sp{gag{-}mos}$ transforming protein, and was found to have a two base deletion. This deletion accomplished the frameshift necessary for v-mos expression, but did not alter MuSVts110 RNA splicing and the expression of p85$\sp{gag{-}mos}.$ Lastly, sequencing of the MuSVts110 DNA from three spontaneous revertants revealed the same G to T transversion in each one. This was the same mutation that was found in the Cd-B2 revertant. These findings provide the first example of mutations resulting from exposure to cadmium and suggest, by the difference in each mutation, the complexity of the mechanism utilized by cadmium to induce DNA damage. ^

AN INVESTIGATION OF VOLATILE ORGANIC AIR CONTAMINANTS IN THE INDOOR MANUFACTURED HOUSING ENVIRONMENT (TEXAS)

Manufactured housing has been found to have substantial levels of formaldehyde in the indoor air. Because mobile homes are more affordable than conventional housing, there has been a large increase in their use in the U.S. This increase in mobile home use has been substantial in the sunbelt regions such as Texas, where high temperatures and humidities may enhance out-gassing of formaldehyde and other volatile organic compounds from construction and furnishing materials and increase any potential health hazards.^ The influences of environmental, architectural and temporal factors on the presence of indoor formaldehyde and other organic compounds were investigated in conjunction with the Texas Indoor Air Quality Study of manufactured housing. A matched pair of mobile homes, one with electric heating and cooking utilities and the other with propane gas utilities, were used for a series of controlled experiments over a fourteen month period from October, 1982 through November, 1983.^ Over this fourteen month period formaldehyde levels decreased approximately 33%. Daily fluctuations of 20% to 40% were observed even with a constant indoor temperature. An increase in indoor temperature of 8(DEGREES)C doubled the measured formaldehyde concentration. Opening windows resulted in decreases of indoor formaldehyde levels of up to 50%. Studies of the impact of propane as a cooking source showed no increase in formaldehyde levels with stove use.^ The presence and concentration of selected volatile organic compounds is influenced greatest by occupancy. Occupants continually open and close windows and doors, vary the operation and settings (temperature) of air control systems, and vary in their selection of furnishings and use of consumer products, which may act as sources of indoor air contaminants. ^