Mechanism of protein synthesis and a high-throughput drug screening assay in mycobacteria

Purpose and goal

1. We have developed a fully reconstituted transcription-translation-folding system or `RTTF system´ based on Escherichia coli. A similar system using components from Mycobacteria smegmatis is half-way through. 2. We have, in collaboration with Umesh Varshney´s group at IISC Bangalore, started biochemical experiments to explore different steps of protein synthesis in Mycobacteria. 3. We have, also succeeded to develop a high-throughput drug screening platform using the E. coli RTTF system and tested several hit compounds from Sanofi-Avantes.

Results and expected effects

1. Strong research collaboration could be established between Suparna Sanyal´s research group at Uppsala University and Professor Umesh Varshney´s group at the Indian Institute of Science, Bangalore, India. 2. Suparna Sanyal started a sole-proprietorship company called `RTTF-Technologies´, which aims to commercialize the RTTF system and the knowledge acquired from it. 3. Sanyal group published three articles and one book-chapter from this project and two manuscripts are under preparation.

Approach and implementation

The project was designed to develop a new, powerful and well-tuned reconstituted system for 1) studying transcription, translation and folding in Mycobacteria, as well as 2) for developing a high through-put screening system for anti-mycobacterial drugs. The development of the bacterial RTTF system has been successfully implemented within the project period. We have made significant progress in biochemical analysis of mycobacterial protein synthesis. Moreover, testing of compounds as inhibitors of protein synthesis has been successfully performed using the RTTF system.