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Sequences upstream from the mouse c-mos oncogene may function as a transcription termination signal
DNA (Mary Ann Liebert, Inc.)
|August 1, 1986
Summary
The upstream mouse sequence (UMS) near the c-mos proto-oncogene contains polyadenylation signals and acts as a transcription terminator. This UMS region inhibits gene expression by terminating RNA transcripts, impacting mos transforming activity.
Area of Science:
- Molecular Biology
- Oncogenes
- Gene Regulation
Background:
- The upstream mouse sequence (UMS) is located near the mouse c-mos proto-oncogene.
- Previous research suggested UMS inhibits mos transforming activity by preventing transcription readthrough.
Purpose of the Study:
- To investigate the mechanism by which the UMS inhibits mos transforming activity.
- To determine if UMS functions as a transcription terminator.
Main Methods:
- Constructed recombinant DNA clone pHTS3MS with UMS downstream of the mos gene.
- Analyzed RNA expression in transformed cells using S1 nuclease analysis.
- Performed nuclear run-on transcription assays.
- Inserted UMS fragments upstream of the herpes simplex virus type 1 thymidine kinase (tk) gene.
Main Results:
- When UMS was placed downstream of the mos gene, it did not inhibit mos transforming activity.
- S1 nuclease analysis revealed UMS provides polyadenylation signals for mos-containing RNA.
- Nuclear run-on transcription demonstrated primary transcripts terminate within UMS.
- A 360-bp UMS fragment inhibited tk transforming activity by 99% and prevented expression.
Conclusions:
- The UMS contains functional polyadenylation signals.
- The UMS acts as a transcription terminator, explaining its inhibitory effect on gene expression.