The integrity of the G2421-C2395 base pair in the ribosomal E-site is crucial for protein synthesis.

Koch, Miriam; Clementi, Nina; Rusca, Nicola; Vögele, Paul; Erlacher, Matthias; Polacek, Norbert (2015). The integrity of the G2421-C2395 base pair in the ribosomal E-site is crucial for protein synthesis. RNA biology, 12(1), pp. 70-81. Taylor and Francis 10.1080/15476286.2015.1017218

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During the elongation cycle of protein biosynthesis, tRNAs traverse through the ribosome by consecutive binding to the 3 ribosomal binding sites (A-, P-, and E- sites). While the ribosomal A- and P-sites have been functionally well characterized in the past, the contribution of the E-site to protein biosynthesis is still poorly understood in molecular terms. Previous studies suggested an important functional interaction of the terminal residue A76 of E-tRNA with the nucleobase of the universally conserved 23S rRNA residue C2394. Using an atomic mutagenesis approach to introduce non-natural nucleoside analogs into the 23S rRNA, we could show that removal of the nucleobase or the ribose 2'-OH at C2394 had no effect on protein synthesis. On the other hand, our data disclose the importance of the highly conserved E-site base pair G2421-C2395 for effective translation. Ribosomes with a disrupted G2421-C2395 base pair are defective in tRNA binding to the E-site. This results in an impaired translation of genuine mRNAs, while homo-polymeric templates are not affected. Cumulatively our data emphasize the importance of E-site tRNA occupancy and in particular the intactness of the 23S rRNA base pair G2421-C2395 for productive protein biosynthesis.

Item Type:

Journal Article (Original Article)

Division/Institute:

08 Faculty of Science > Departement of Chemistry and Biochemistry

UniBE Contributor:

Koch, Miriam; Erlacher, Matthias and Polacek, Norbert

Subjects:

500 Science > 570 Life sciences; biology
500 Science > 540 Chemistry

ISSN:

1555-8584

Publisher:

Taylor and Francis

Language:

English

Submitter:

Christina Schüpbach

Date Deposited:

01 May 2015 10:56

Last Modified:

08 Sep 2017 13:21

Publisher DOI:

10.1080/15476286.2015.1017218

PubMed ID:

25826414

Uncontrolled Keywords:

23S rRNA; E-site E-tRNA; atomic mutagenesis; ribosome translation

BORIS DOI:

10.7892/boris.67761

URI:

https://boris.unibe.ch/id/eprint/67761

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