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.
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