BIOCHEMICAL ANALYSIS OF COMPLEX FORMATION BETWEEN RNA POLYMERASE AND RIBOSOME

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Authors	HIRIYUR NAGARAJ Pradeep HASSAN Ahmed Adel I. H. BRABENCOVÁ Sylva SLÁMOVÁ Lucie DEMO Gabriel
Year of publication	2022
Type	Conference abstract
MU Faculty or unit	Central European Institute of Technology
Citation
Description	Transcription of messenger RNAs and translation of proteins, mechanisms necessary for gene expression, can be physically coupled and coordinated in some bacterial species1,2. The interaction between the leading ribosome and RNA polymerase (RNAP) forms a transcription-translation coupling complex (CTT)2–4 modulated by transcription elongation factor NusG3,4. The leading ribosome also regulates the propagation of the RNAP during transcription1 and protects the RNAP from backtracking. Moreover, the transcription pausing and possible premature termination can be modulated by cis-acting RNA structures (riboswitches) which may play a crucial role in formation of coupled transcription-translation. Here, we investigate the coordination of translation initiation and elongation with RNAP pausing by forming CTT complexes in sucrose gradients. For translation initiation we use specific riboswitch to form and stabilize the initiation complex during early steps of CTT formation. For translation elongation we load the aminoacylated-tRNA into the CTT in the form of ternary complex.
Related projects:	National Institute of Virology and Bacteriology