Comparative toxic potential of market formulation of two organophosphate pesticides in transgenic Drosophila melanogaster (hsp70-lacZ).

This study investigated the working hypothesis that two widely used organophosphate pesticides; Nuvan and Dimecron, exert toxic effects in Drosophila. Transgenic D. melanogaster (hsp70-lacZ) was used as a model for assaying stress gene expression and AchE activity as an endpoint for toxicity and also to evaluate whether stress gene expression is sufficient to protect against toxic insult of the chemicals and to prevent tissue damage. The study was extended to investigate the effect of the pesticides on the life cycle and reproduction of the organism. The study showed that Nuvan affected emergence of the exposed flies more drastically than Dimecron and the effect was lethal at the highest tested concentration (0.075 ppm). While Nuvan at 0.0075 and 0.015 ppm concentrations affected reproduction of the flies significantly, the effect of Dimecron was significant only at 0.015 and 0.075 ppm. Nuvan-exposed third-instar larvae exhibited a 1.2-fold to 1.5-fold greater hsp70 expression compared to Dimecron at concentrations ranging from 0.0075 to 0.075 ppm following 12 and 18 h exposure. While maximum expression of hsp70 was observed in Nuvan-exposed third-instar larval tissues following 18 h exposure at 0.075 ppm, Dimecron at the same dietary concentration induced a maximum expression of hsp70 following 24 h exposure. Further, concomitant with a significant induction of hsp70, significant inhibition of AchE was observed following chemical exposure and temperature shock. Concurrent with a significant decline in hsp70 expression in Nuvan-exposed larvae after 48 h at 0.075 ppm, tissue damage was evident. Dimecron-exposed larvae exhibited a plateau in hsp70 induction even after 48 h exposure and moderate tissue damage was observed in these larvae. The present study suggests that Nuvan is more cytotoxic than Dimecron in transgenic Drosophila melanogaster.

Characterization of the DNA binding and transcriptional activation domains of the erg protein.

erg, an ets related gene encodes a sequence specific DNA binding transcriptional activator protein. We have identified four functional domains of erg protein that are responsible for DNA binding, transcriptional activation and negative regulation of transcriptional activation. Deletion analysis revealed that the 3'-ets domain of the erg protein is sufficient for DNA binding activity. Analysis of these deletion mutants also revealed the presence of two autonomous transcriptional activation domains, one at the amino and the other at the carboxyterminal region. This aminoterminal transcriptional activator domain (5'-ets domain) is conserved in six of the ets genes suggesting that it (ETA, ets Transcriptional Activation domain) may contribute to a common function among these genes. The transcriptional activation function of the carboxy terminal transcriptional activation domain (CTA) was inhibited by the presence of a Negative Regulatory Transcriptional activation domain (NRT), which is located at the amino terminal region of erg DNA binding domain. These results may help in understanding the structure/function relationship of other erg/ets related proteins.

The interaction of a c-Jun/Fos related protein factor with the U3 sequences of the mouse mammary tumor virus LTR.

Using polyacrylamide gel mobility shift assay we have detected a nuclear factor (NF-S) in a mouse mammary tumor cell line (GR) that binds to an upstream sequence domain (-766 to -737) near the 5'-end of the mouse mammary tumor virus (MMTV) long terminal repeat (LTR). Antibodies to the products of the Jun and Fos oncogenes interfered with the binding potential of this factor, indicating that the factor shares antigenic determinants with c-Jun/AP-1. In vitro translated c-Jun and c-Fos were also found to bind to the NF-S binding domain consisting of the sequence TGA(A/G)TCA that are known to be recognized by c-Jun/AP-1. Our results raise the possibility that the MMTV-LTR sequence element-766 to -737 by interacting with a Jun/Fos related protein play a role in MMTV transcription and/or the activation of int protooncogenes that are associated with murine mammary tumorigenesis.