The ribosome modulates the structural dynamics of the conserved GTPase HflX and triggers tight nucleotide binding

Biochimie
Jeffrey J FischerHans-Joachim Wieden

Abstract

The universally conserved GTPase HflX is a putative translation factor whose GTPase activity is stimulated by the 70S ribosome as well as the 50S but not the 30S ribosomal subunit. However, the details and mechanisms governing this interaction are only poorly understood. In an effort to further elucidate the functional mechanism of HflX, we examined its interaction with the 70S ribosome, the two ribosomal subunits (50S and 30S), as well as its ability to interact with guanine nucleotides in the respective ribosomal complexes using a highly purified in vitro system. Binding studies reported here demonstrate that HflX not only interacts with 50S and 70S particles, but also with the 30S subunit, independent of the nucleotide-bound state. A detailed pre-steady-state kinetic analysis of HflX interacting with a non-hydrolyzable analog of mant-GTP, coupled with an enzymatic probing assay utilizing limited trypsinolysis, reveal that HflX·GTP exists in a structurally distinct 50S- and 70S-bound form that stabilizes GTP binding up to 70 000-fold and that may represent the "GTPase-activated" state. This activation is likely required for efficient GTP-hydrolysis, and may be similar to that observed in elongation factor G. Results reported ...Continue Reading

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Citations

Jan 7, 2016·Nucleic Acids Research·Mackenzie L CoathamHans-Joachim Wieden
Oct 13, 2015·Nature Structural & Molecular Biology·Yanqing ZhangNing Gao
Apr 14, 2016·BMC Microbiology·Carolina A P T da SilvaMarilis V Marques
Sep 22, 2012·RNA Biology·Marion BeckerHans-Joachim Wieden
May 21, 2015·The Journal of Biological Chemistry·Adrián García-MárquezNuria Sánchez-Puig
Feb 26, 2019·Microbial Biotechnology·Grégory BoëlAntoine Danchin
Nov 19, 2018·Current Genetics·Krishnamoorthi SrinivasanJayati Sengupta
Mar 17, 2021·Proceedings of the National Academy of Sciences of the United States of America·Jie Yin Grace NganSylvie Alonso

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