Effects of antibiotics on gene expression in D. radiodurans
EMSL Project ID
2234
Abstract
Main objective of this proposal is the characterization of the changes in gene expression in Deinococcus radiodurans due to the presence of known antibiotics with defined mode of action and demonstrated activity against D. radiodurans strain. All standard antibiotics tested have well-defined mechanisms such as dihydrofolate reductase inhibition (trimethoprim), protein biosynthesis inhibition (oxytetracycline, clindamycin, macrolides), stabilization of cleavable complex with topoisomerase and DNA (quinolones). This proposal is particularly attractive in the light of the work already done at EMSL, i.e., development of an approach that uses global proteolytic protein digests and highly sensitive FTICR mass spectrometry measurements to define the proteins actually expressed by D. radiodurans. The high FTICR mass measurement accuracy (MMA) enabled many peptide masses to be uniquely attributed to D. radiodurans proteins. MS/MS have then been used for confirmation of these assignments, and a database of such accurate mass tags (AMTs) was then constructed. This methodology enabled conformation of almost half of the proteins predicted by D. radiodurans genome annotation. Its key advantage is that subsequent high throughput quantitative measurements of changes in gene expression (as the one proposed here) are based upon predetermined AMTs with the use of stable isotope labeling and the high MMA achievable by FTICR MS. We propose to use existing data and methodology to characterize proteome-wide effects of the known antibiotics on D. radiodurans.
Project Details
Project type
Exploratory Research
Start Date
2001-10-09
End Date
2001-12-31
Status
Closed
Released Data Link
Team
Principal Investigator