Home LiteratureArticle Details
PMID: 22992746 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, U.S. Gov't, Non-P.H.S.

The selenocysteine-specific elongation factor contains a novel and multi-functional domain.

The Journal of biological chemistry ·Vol. 287 ·No. 46 ·2012-11-09 ·Pages 38936-45

Gonzalez-Flores JN, Gupta N, DeMong LW, Copeland PR

Abstract

The selenocysteine (Sec)-specific eukaryotic elongation factor (eEFSec) delivers the aminoacylated selenocysteine-tRNA (Sec-tRNA(Sec)) to the ribosome and suppresses UGA codons that are upstream of Sec insertion sequence (SECIS) elements bound by SECIS-binding protein 2 (SBP2). Multiple studies have highlighted the importance of SBP2 forming a complex with the SECIS element, but it is not clear how this regulates eEFSec during Sec incorporation. Compared with the canonical elongation factor eEF1A, eEFSec has a unique C-terminal extension called Domain IV. To understand the role of Domain IV in Sec incorporation, we examined a series of mutant proteins for all of the known molecular functions for eEFSec: GTP hydrolysis, Sec-tRNA(Sec) binding, and SBP2/SECIS binding. In addition, wild-type and mutant versions of eEFSec were analyzed for Sec incorporation activity in a novel eEFSec-dependent translation extract. We have found that Domain IV is essential for both tRNA and SBP2 binding as well as regulating GTPase activity. We propose a model where the SBP2/SECIS complex activates eEFSec by directing functional interactions between Domain IV and the ribosome to promote Sec-tRNA(Sec) binding and accommodation into the ribosomal A-site.

MeSH Terms
Amino Acid Sequence Animals Binding Sites Codon Codon, Terminator Cross-Linking Reagents/chemistry Guanosine Triphosphate/chemistry Hydrolysis Mice Molecular Sequence Data Peptide Elongation Factors/chemistry,metabolism Protein Biosynthesis Protein Structure, Tertiary RNA, Transfer, Amino Acyl/chemistry RNA-Binding Proteins/metabolism Rats Recombinant Proteins/chemistry Ribosomes/chemistry Selenocysteine/chemistry Sequence Homology, Amino Acid
Chemicals
Codon Codon, Terminator Cross-Linking Reagents EFsec protein, mouse Peptide Elongation Factors RNA, Transfer, Amino Acyl RNA-Binding Proteins Recombinant Proteins SECIS-binding protein 2, mouse Secisbp2 protein, rat selenocysteinyl-tRNA Selenocysteine Guanosine Triphosphate
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Gonzalez-Flores Jonathan N
Department of Biochemistry and Molecular Biology, Robert Wood Johnson Medical School, University of Medicine and Dentistry of New Jersey, Piscataway, New Jersey 08854, USA.
Gupta Nirupama
DeMong Louise W
Copeland Paul R
References (43)
43 references, click to expand
  1. Domain structure of the prokaryotic selenocysteine-specific elongation factor SelB.
    J Mol Biol. 1996 Oct 4;262(4):413-20 PMID: 8893853
  2. Position of eukaryotic initiation factor eIF5B on the 80S ribosome mapped by directed hydroxyl radical probing.
    EMBO J. 2007 Jul 11;26(13):3109-23 PMID: 17568775
  3. Size matters: a view of selenocysteine incorporation from the ribosome.
    Cell Mol Life Sci. 2006 Jan;63(1):73-81 PMID: 16416259
  4. Characterization of mSelB, a novel mammalian elongation factor for selenoprotein translation.
    EMBO J. 2000 Sep 1;19(17):4796-805 PMID: 10970870
  5. An improved definition of the RNA-binding specificity of SECIS-binding protein 2, an essential component of the selenocysteine incorporation machinery.
    Nucleic Acids Res. 2007;35(6):1868-84 PMID: 17332014
  6. Structural basis for mRNA recognition by elongation factor SelB.
    Nat Struct Mol Biol. 2005 Feb;12(2):198-203 PMID: 15665870
  7. Selenocysteine tRNA-specific elongation factor SelB is a structural chimaera of elongation and initiation factors.
    EMBO J. 2005 Jan 12;24(1):11-22 PMID: 15616587
  8. Structural insight into a molecular switch in tandem winged-helix motifs from elongation factor SelB.
    J Mol Biol. 2007 Jul 20;370(4):728-41 PMID: 17537456
  9. Selenoprotein expression is regulated at multiple levels in prostate cells.
    Cell Res. 2006 Dec;16(12):940-8 PMID: 17160069
  10. Decoding apparatus for eukaryotic selenocysteine insertion.
    EMBO Rep. 2000 Aug;1(2):158-63 PMID: 11265756
  11. Functional characterization of the eukaryotic SECIS elements which direct selenocysteine insertion at UGA codons.
    EMBO J. 1993 Aug;12(8):3315-22 PMID: 8344267
  12. Structure of prokaryotic SECIS mRNA hairpin and its interaction with elongation factor SelB.
    J Mol Biol. 2002 Nov 15;324(1):137-50 PMID: 12421564
  13. Coupled tRNA(Sec)-dependent assembly of the selenocysteine decoding apparatus.
    Mol Cell. 2003 Mar;11(3):773-81 PMID: 12667458
  14. Initiation factor IF 2 binds to the alpha-sarcin loop and helix 89 of Escherichia coli 23S ribosomal RNA.
    RNA. 2001 Aug;7(8):1173-9 PMID: 11497435
  15. Selenocysteine insertion sequence (SECIS)-binding protein 2 alters conformational dynamics of residues involved in tRNA accommodation in 80 S ribosomes.
    J Biol Chem. 2012 Mar 23;287(13):10664-10673 PMID: 22308032
  16. Thermodynamic and kinetic framework of selenocysteyl-tRNASec recognition by elongation factor SelB.
    J Biol Chem. 2010 Jan 29;285(5):3014-20 PMID: 19940162
  17. Cell-free protein synthesis system prepared from insect cells by freeze-thawing.
    Biotechnol Prog. 2006 Nov-Dec;22(6):1570-7 PMID: 17137303
  18. A novel protein domain induces high affinity selenocysteine insertion sequence binding and elongation factor recruitment.
    J Biol Chem. 2008 Dec 12;283(50):35129-39 PMID: 18948268
  19. A novel RNA structural motif in the selenocysteine insertion element of eukaryotic selenoprotein mRNAs.
    RNA. 1996 Apr;2(4):367-79 PMID: 8634917
  20. Class-1 release factor eRF1 promotes GTP binding by class-2 release factor eRF3.
    Biochimie. 2006 Jul;88(7):747-57 PMID: 16797113
  21. Crystal structure of an mRNA-binding fragment of Moorella thermoacetica elongation factor SelB.
    EMBO J. 2002 Aug 1;21(15):4145-53 PMID: 12145214
  22. Ribosome-induced changes in elongation factor Tu conformation control GTP hydrolysis.
    Proc Natl Acad Sci U S A. 2009 Jan 27;106(4):1063-8 PMID: 19122150
  23. Structural studies of elongation and release factors.
    Cell Mol Life Sci. 2008 May;65(9):1335-46 PMID: 18213444
  24. Mammalian selenoprotein in which selenocysteine (Sec) incorporation is supported by a new form of Sec insertion sequence element.
    Mol Cell Biol. 2002 Mar;22(5):1402-11 PMID: 11839807
  25. Efficiency of mammalian selenocysteine incorporation.
    J Biol Chem. 2004 Sep 3;279(36):37852-9 PMID: 15229221
  26. The L7Ae RNA binding motif is a multifunctional domain required for the ribosome-dependent Sec incorporation activity of Sec insertion sequence binding protein 2.
    Mol Cell Biol. 2007 Sep;27(18):6350-60 PMID: 17636016
  27. The many applications of acid urea polyacrylamide gel electrophoresis to studies of tRNAs and aminoacyl-tRNA synthetases.
    Methods. 2008 Feb;44(2):129-38 PMID: 18241794
  28. Structural basis for dynamic interdomain movement and RNA recognition of the selenocysteine-specific elongation factor SelB.
    Structure. 2007 May;15(5):577-86 PMID: 17502103
  29. Novel structural determinants in human SECIS elements modulate the translational recoding of UGA as selenocysteine.
    Nucleic Acids Res. 2009 Sep;37(17):5868-80 PMID: 19651878
  30. Selenocysteine inserting RNA elements modulate GTP hydrolysis of elongation factor SelB.
    Biochemistry. 1998 Jan 20;37(3):885-90 PMID: 9454578
  31. How can elongation factors EF-G and EF-Tu discriminate the functional state of the ribosome using the same binding site?
    FEBS Lett. 2005 Oct 24;579(25):5439-42 PMID: 16213500
  32. A novel RNA binding protein, SBP2, is required for the translation of mammalian selenoprotein mRNAs.
    EMBO J. 2000 Jan 17;19(2):306-14 PMID: 10637234
  33. Crystal structure of human selenocysteine tRNA.
    Nucleic Acids Res. 2009 Oct;37(18):6259-68 PMID: 19692584
  34. The selenocysteine incorporation machinery: interactions between the SECIS RNA and the SECIS-binding protein SBP2.
    RNA. 2001 Oct;7(10):1442-53 PMID: 11680849
  35. Structural rearrangements of the ribosome at the tRNA proofreading step.
    Nat Struct Mol Biol. 2010 Sep;17(9):1072-8 PMID: 20694005
  36. Interaction of translation factor SELB with the formate dehydrogenase H selenopolypeptide mRNA.
    Proc Natl Acad Sci U S A. 1993 May 1;90(9):4181-5 PMID: 8483932
  37. Thermodynamics of the GTP-GDP-operated conformational switch of selenocysteine-specific translation factor SelB.
    J Biol Chem. 2012 Aug 10;287(33):27906-12 PMID: 22740700
  38. [Interaction of 23S ribosomal RNA helices 89 and 91 of Escherichia coli contributes to the activity of IF2 but is insignificant for elongation factors functioning].
    Mol Biol (Mosk). 2007 Nov-Dec;41(6):1031-41 PMID: 18318122
  39. Evolutionary history of selenocysteine incorporation from the perspective of SECIS binding proteins.
    BMC Evol Biol. 2009 Sep 10;9:229 PMID: 19744324
  40. Characterization of the SECIS binding protein 2 complex required for the co-translational insertion of selenocysteine in mammals.
    Nucleic Acids Res. 2005 Sep 09;33(16):5172-80 PMID: 16155186
  41. The mechanism for activation of GTP hydrolysis on the ribosome.
    Science. 2010 Nov 5;330(6005):835-838 PMID: 21051640
  42. SECISaln, a web-based tool for the creation of structure-based alignments of eukaryotic SECIS elements.
    Bioinformatics. 2009 Mar 1;25(5):674-5 PMID: 19179357
  43. Protection against reactive oxygen species by selenoproteins.
    Biochim Biophys Acta. 2009 Nov;1790(11):1478-85 PMID: 19268692
Article Info
Journal
The Journal of biological chemistry
Abbr.
J Biol Chem
ISSN
1083-351X
DOI
10.1074/jbc.M112.415463
Published
2012-11-09
Epub
2012-00-19
Pages
38936-45
Language
English
Region
United States
NLM ID
2985121R
PMCID
PMC3493934
Subset
IM
Grants
NIGMS NIH HHS · R01 GM077073 · United States
NIGMS NIH HHS · T32 GM008339 · United States
NIGMS NIH HHS · GM077073 · United States
NIGMS NIH HHS · 2T32GM008339-21 · United States
External Links
PubMed source DOI: 10.1074/jbc.M112.415463 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2