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PMID: 17636016 Published · ppublish English Journal Article Research Support, N.I.H., Extramural

The L7Ae RNA binding motif is a multifunctional domain required for the ribosome-dependent Sec incorporation activity of Sec insertion sequence binding protein 2.

Molecular and cellular biology ·Vol. 27 ·No. 18 ·2007-09-00 ·Pages 6350-60

Caban K, Kinzy SA, Copeland PR

Abstract

The decoding of specific UGA codons as selenocysteine is specified by the Sec insertion sequence (SECIS) element. Additionally, Sec-tRNA([Ser]Sec) and the dedicated Sec-specific elongation factor eEFSec are required but not sufficient for nonsense suppression. SECIS binding protein 2 (SBP2) is also essential for Sec incorporation, but its precise role is unknown. In addition to binding the SECIS element, SBP2 binds stably and quantitatively to ribosomes. To determine the function of the SBP2-ribosome interaction, conserved amino acids throughout the SBP2 L7Ae RNA binding motif were mutated to alanine in clusters of five. Mutant proteins were analyzed for ribosome binding, SECIS element binding, and Sec incorporation activity, allowing us to identify two distinct but interdependent sites within the L7Ae motif: (i) a core L7Ae motif required for SECIS binding and ribosome binding and (ii) an auxiliary motif involved in physical and functional interactions with the ribosome. Structural modeling of SBP2 based on the 15.5-kDa protein-U4 snRNA complex strongly supports a two-site model for L7Ae domain function within SBP2. These results provide evidence that the SBP2-ribosome interaction is essential for Sec incorporation.

MeSH Terms

Alanine/metabolism Amino Acid Motifs Amino Acid Sequence Amino Acid Substitution Animals Conserved Sequence Male Models, Molecular Molecular Sequence Data Protein Structure, Tertiary RNA-Binding Proteins/chemistry,genetics,physiology Rats Ribosomes/metabolism Selenocysteine/metabolism Sequence Homology, Amino Acid

Chemicals

RNA-Binding Proteins Secisbp2 protein, rat Selenocysteine Alanine

Authors & Affiliations

3 authors, click to expand affiliations / ORCID

Caban Kelvin

Department of Molecular Genetics, Microbiology and Immunology, UMDNJ-Robert Wood Johnson Medical School, 675 Hoes Lane, Piscataway, NJ 08854, USA.

Kinzy Scott A

Copeland Paul R

References (31)

31 references, click to expand

Yeast as a sensor of factors affecting the accuracy of protein synthesis.
Cell Mol Life Sci. 2003 Oct;60(10):2115-30 PMID: 14618259
The SBP2 and 15.5 kD/Snu13p proteins share the same RNA binding domain: identification of SBP2 amino acids important to SECIS RNA binding.
RNA. 2002 Oct;8(10):1308-18 PMID: 12403468
The PredictProtein server.
Nucleic Acids Res. 2004 Jul 1;32(Web Server issue):W321-6 PMID: 15215403
Joint X-ray and NMR refinement of the yeast L30e-mRNA complex.
Structure. 2004 Jul;12(7):1165-76 PMID: 15242593
Ribosomal RNA kink-turn motif--a flexible molecular hinge.
J Biomol Struct Dyn. 2004 Oct;22(2):183-94 PMID: 15317479
Efficiency of mammalian selenocysteine incorporation.
J Biol Chem. 2004 Sep 3;279(36):37852-9 PMID: 15229221
Selenocysteine tRNA[Ser]Sec gene is ubiquitous within the animal kingdom.
Mol Cell Biol. 1990 May;10(5):1940-9 PMID: 2139169
The hnRNP F protein: unique primary structure, nucleic acid-binding properties, and subcellular localization.
Nucleic Acids Res. 1994 Mar 25;22(6):1059-67 PMID: 7512260
A novel RNA-binding motif in omnipotent suppressors of translation termination, ribosomal proteins and a ribosome modification enzyme?
Nucleic Acids Res. 1994 Jun 11;22(11):2166-7 PMID: 7518079
UGA suppression by a mutant RNA of the large ribosomal subunit.
Proc Natl Acad Sci U S A. 1995 Dec 19;92(26):12309-13 PMID: 8618891
Selenocysteine incorporation in eukaryotes: insights into mechanism and efficiency from sequence, structure, and spacing proximity studies of the type 1 deiodinase SECIS element.
RNA. 1996 Feb;2(2):171-82 PMID: 8601283
Interaction between the human nuclear cap-binding protein complex and hnRNP F.
Mol Cell Biol. 1997 May;17(5):2587-97 PMID: 9111328
hnRNP H is a component of a splicing enhancer complex that activates a c-src alternative exon in neuronal cells.
Mol Cell Biol. 1999 Jan;19(1):69-77 PMID: 9858532
Purification, redox sensitivity, and RNA binding properties of SECIS-binding protein 2, a protein involved in selenoprotein biosynthesis.
J Biol Chem. 1999 Sep 3;274(36):25447-54 PMID: 10464275
Discrimination between defects in elongation fidelity and termination efficiency provides mechanistic insights into translational readthrough.
J Mol Biol. 2005 May 13;348(4):801-15 PMID: 15843014
Ribosomal protein L30 is a component of the UGA-selenocysteine recoding machinery in eukaryotes.
Nat Struct Mol Biol. 2005 May;12(5):408-16 PMID: 15821744
Localization and dynamic behavior of ribosomal protein L30e.
Nat Struct Mol Biol. 2005 May;12(5):467-8 PMID: 15864315
The crystal structure of the Methanocaldococcus jannaschii multifunctional L7Ae RNA-binding protein reveals an induced-fit interaction with the box C/D RNAs.
Biochemistry. 2005 Jul 19;44(28):9657-72 PMID: 16008351
Characterization of the SECIS binding protein 2 complex required for the co-translational insertion of selenocysteine in mammals.
Nucleic Acids Res. 2005;33(16):5172-80 PMID: 16155186
Aminoglycosides decrease glutathione peroxidase-1 activity by interfering with selenocysteine incorporation.
J Biol Chem. 2006 Feb 10;281(6):3382-8 PMID: 16354666
RNA kink-turns as molecular elbows: hydration, cation binding, and large-scale dynamics.
Structure. 2006 May;14(5):825-35 PMID: 16698544
Aminoglycoside antibiotics mediate context-dependent suppression of termination codons in a mammalian translation system.
RNA. 2000 Jul;6(7):1044-55 PMID: 10917599
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
The redox state of SECIS binding protein 2 controls its localization and selenocysteine incorporation function.
Mol Cell Biol. 2006 Jul;26(13):4895-910 PMID: 16782878
The complete atomic structure of the large ribosomal subunit at 2.4 A resolution.
Science. 2000 Aug 11;289(5481):905-20 PMID: 10937989
Characterization of mSelB, a novel mammalian elongation factor for selenoprotein translation.
EMBO J. 2000 Sep 1;19(17):4796-805 PMID: 10970870
Crystal structure of the spliceosomal 15.5kD protein bound to a U4 snRNA fragment.
Mol Cell. 2000 Dec;6(6):1331-42 PMID: 11163207
Insight into mammalian selenocysteine insertion: domain structure and ribosome binding properties of Sec insertion sequence binding protein 2.
Mol Cell Biol. 2001 Mar;21(5):1491-8 PMID: 11238886
Decoding apparatus for eukaryotic selenocysteine insertion.
EMBO Rep. 2000 Aug;1(2):158-63 PMID: 11265756
The kink-turn: a new RNA secondary structure motif.
EMBO J. 2001 Aug 1;20(15):4214-21 PMID: 11483524
Structure of protein L7Ae bound to a K-turn derived from an archaeal box H/ACA sRNA at 1.8 A resolution.
Structure. 2004 May;12(5):893-903 PMID: 15130481

Article Info

Journal

Molecular and cellular biology

Abbr.

Mol Cell Biol

ISSN

0270-7306

DOI

10.1128/MCB.00632-07

Published

2007-09-00

Epub

2007-00-16

Pages

6350-60

Language

English

Region

United States

NLM ID

8109087

PMCID

PMC2099609

Subset

Grants

NIGMS NIH HHS · R01 GM068077 · United States

NIGMS NIH HHS · R25 GM066338 · United States

NIGMS NIH HHS · GM068077 · United States

NIGMS NIH HHS · GM66338 · United States

External Links

PubMed source DOI: 10.1128/MCB.00632-07 PMC Full Text View journal details

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The L7Ae RNA binding motif is a multifunctional domain required for the ribosome-dependent Sec incorporation activity of Sec insertion sequence binding protein 2.

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