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PMID: 22308032 Published · ppublish English Journal Article Research Support, N.I.H., Extramural

Selenocysteine insertion sequence (SECIS)-binding protein 2 alters conformational dynamics of residues involved in tRNA accommodation in 80 S ribosomes.

The Journal of biological chemistry ·Vol. 287 ·No. 13 ·2012-03-23 ·Pages 10664-10673

Caban K, Copeland PR

Abstract

Sec-tRNA(Sec) is site-specifically delivered at defined UGA codons in selenoprotein mRNAs. This recoding event is specified by the selenocysteine insertion sequence (SECIS) element and requires the selenocysteine (Sec)-specific elongation factor, eEFSec, and the SECIS binding protein, SBP2. Sec-tRNA(Sec) is delivered to the ribosome by eEFSec-GTP, but this ternary complex is not sufficient for Sec incorporation, indicating that its access to the ribosomal A-site is regulated. SBP2 stably associates with ribosomes, and mutagenic analysis indicates that this interaction is essential for Sec incorporation. However, the ribosomal function of SBP2 has not been elucidated. To shed light on the functional relevance of the SBP2-ribosome interaction, we screened the functional centers of the 28 S rRNA in translationally competent 80 S ribosomes using selective 2'-hydroxyl acylation analyzed by primer extension (SHAPE). We demonstrate that SBP2 specifically alters the reactivity of specific residues in Helix 89 (H89) and expansion segment 31 (ES31). These results are indicative of a conformational change in response to SBP2 binding. Based on the known functions of H89 during translation, we propose that SBP2 allows Sec incorporation by either promoting Sec-tRNA(Sec) accommodation into the peptidyltransferase center and/or by stimulating the ribosome-dependent GTPase activity of eEFSec.

MeSH Terms
Animals GTP Phosphohydrolases/chemistry,genetics,metabolism Humans Mice Mutagenesis Nucleic Acid Conformation Peptidyl Transferases/chemistry,genetics,metabolism Protein Biosynthesis/physiology RNA, Ribosomal, 28S/chemistry,genetics,metabolism RNA, Transfer, Amino Acyl/chemistry,genetics,metabolism RNA-Binding Proteins/chemistry,genetics,metabolism Ribosomes/chemistry,genetics,metabolism
Chemicals
RNA, Ribosomal, 28S RNA, Transfer, Amino Acyl RNA-Binding Proteins SECISBP2 protein, human selenocysteinyl-tRNA Peptidyl Transferases GTP Phosphohydrolases
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Caban Kelvin
Department of Molecular Genetics, Microbiology, and Immunology, University of Medicine and Dentistry of New Jersey, Robert Wood Johnson Medical School, Piscataway, New Jersey 08854.
Copeland Paul R
Department of Molecular Genetics, Microbiology, and Immunology, University of Medicine and Dentistry of New Jersey, Robert Wood Johnson Medical School, Piscataway, New Jersey 08854. Electronic address: paul.copeland@umdnj.edu.
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Article Info
Journal
The Journal of biological chemistry
Abbr.
J Biol Chem
ISSN
1083-351X
DOI
10.1074/jbc.M111.320929
Email
paul.copeland@umdnj.edu
Published
2012-03-23
Epub
2012-00-03
Pages
10664-10673
Language
English
Region
United States
NLM ID
2985121R
PMCID
PMC3323001
Subset
IM
Grants
NIGMS NIH HHS · R01 GM094833 · United States
NIGMS NIH HHS · GM077073 · United States
NIGMS NIH HHS · R01 GM077073 · United States
NIGMS NIH HHS · F31GM081920 · United States
NIGMS NIH HHS · F31 GM081920 · United States
NIGMS NIH HHS · GM094833 · United States
External Links
PubMed source DOI: 10.1074/jbc.M111.320929 PMC Full Text View journal details
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