Archaeal ribosomal stalk protein interacts with translation factors in a nucleotide-independent manner via its conserved C terminus.

Proceedings of the National Academy of Sciences of the United States of America
Naoko NomuraToshio Uchiumi

Abstract

Protein synthesis on the ribosome requires translational GTPase factors to bind to the ribosome in the GTP-bound form, take individual actions that are coupled with GTP hydrolysis, and dissociate, usually in the GDP-bound form. The multiple copies of the flexible ribosomal stalk protein play an important role in these processes. Using biochemical approaches and the stalk protein from a hyperthermophilic archaeon, Pyrococcus horikoshii, we here provide evidence that the conserved C terminus of the stalk protein aP1 binds directly to domain I of the elongation factor aEF-2, irrespective of whether aEF-2 is bound to GTP or GDP. Site-directed mutagenesis revealed that four hydrophobic amino acids at the C terminus of aP1, Leu-100, 103, 106, and Phe-107, are crucial for the direct binding. P1 was also found to bind to the initiation factor aIF5B, as well as aEF-1α, but not aIF2γ, via its C terminus. Moreover, analytical ultracentrifugation and gel mobility shift analyses showed that a heptameric complex of aP1 and aP0, aP0(aP1)(2)(aP1)(2)(aP1)(2), can bind multiple aEF-2 molecules simultaneously, which suggests that individual copies of the stalk protein are accessible to the factor. The functional significance of the C terminus of ...Continue Reading

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Citations

Jan 30, 2014·Biochemistry and Cell Biology = Biochimie Et Biologie Cellulaire·Miaoqing HuAihua Liang
Oct 24, 2015·The International Journal of Biochemistry & Cell Biology·Barbara Michalec-WawiorkaMarek Tchorzewski
Sep 19, 2016·Toxicology and Applied Pharmacology·Amanda E JetztWendie S Cohick
Jan 4, 2017·Biochemical and Biophysical Research Communications·Takayoshi HondaToshio Uchiumi
Jun 14, 2017·Molecular and Cellular Biology·Leszek WawiórkaMarek Tchórzewski
Sep 18, 2020·Nucleic Acids Research·Anna Constance VindSimon Bekker-Jensen
Jun 3, 2015·Genes to Cells : Devoted to Molecular & Cellular Mechanisms·Hirotatsu ImaiToshio Uchiumi
Jul 17, 2018·Nucleic Acids Research·Hirotatsu ImaiToshio Uchiumi
Mar 7, 2017·Biochemistry. Biokhimii︠a︡·I V MitroshinA G Gabdulkhakov
Sep 4, 2020·The Journal of Biological Chemistry·Michael J RudolphXiao-Ping Li
Dec 9, 2020·Proceedings of the National Academy of Sciences of the United States of America·Hirotatsu ImaiNoriyuki Kodera

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