HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

J. Biol. Chem., Vol. 279, Issue 36, 37852-37859, September 3, 2004

This Article Full Text Full Text (PDF) All Versions of this Article: 279/36/37852    most recent M404639200v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Mehta, A. Articles by Copeland, P. R. Search for Related Content PubMed PubMed Citation Articles by Mehta, A. Articles by Copeland, P. R. Social Bookmarking                      What's this?

Efficiency of Mammalian Selenocysteine Incorporation^*

Anupama Mehta, Cheryl M. Rebsch, Scott A. Kinzy, Julia E. Fletcher¶, and Paul R. Copeland||

From the Department of Molecular Genetics, Microbiology and Immunology, University of Medicine and Dentistry of New Jersey Robert Wood Johnson Medical School, Piscataway, New Jersey 08854 and the Department of Cell Biology, Lerner Research Institute, Cleveland Clinic Foundation, Cleveland, Ohio 44195

Five components have thus far been identified that are necessary for the incorporation of selenocysteine (Sec) into 25 mammalian proteins. Two of these are cis sequences, a SECIS element in the 3'-untranslated region and a Sec codon (UGA) in the coding region. The three known trans-acting factors are a Sec-specific translation elongation factor (eEFSec), the Sec-tRNA^Sec, and a SECIS-binding protein, SBP2. Here we describe a system in which the efficiency of Sec incorporation was determined quantitatively both in vitro and in transfected cells, and in which the contribution of each of the known factors is examined. The efficiency of Sec incorporation into a luciferase reporter system in vitro is maximally 5-8%, which is 6-10 times higher than that in transfected rat hepatoma cells, McArdle 7777. In contrast, the efficiency of Sec incorporation into selenoprotein P in vitro is 40%, suggesting that as yet unidentified cis-elements may regulate differential selenoprotein expression. In addition, we have found that SBP2 is the only limiting factor in rabbit reticulocyte lysate but not in transfected rat hepatoma cells where SBP2 is found to be mostly if not entirely cytoplasmic despite having a strong putative nuclear localization signal. The significance of these findings with regard to the function of known Sec incorporation factors is discussed.

Received for publication, April 26, 2004 , and in revised form, June 18, 2004.

^* This work was supported by Public Health Service Grant GM068077 from the National Institutes of Health. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

^¶ Current address: Invitrogen Corporation, Research and Development, Proteomics Platforms, Carlsbad, CA 92008.

^|| To whom correspondence should be addressed: Dept. of Molecular Genetics, Microbiology and Immunology, UMDNJ Robert Wood Johnson Medical School, 675 Hoes Lane, Rm. 728, Piscataway, NJ 08854. Tel.: 732-235-4670; Fax: 732-235-5223; E-mail: paul.copeland{at}umdnj.edu.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				M. Gupta and P. R. Copeland Functional Analysis of the Interplay between Translation Termination, Selenocysteine Codon Context, and Selenocysteine Insertion Sequence-binding Protein 2 J. Biol. Chem., December 21, 2007; 282(51): 36797 - 36807. [Abstract] [Full Text] [PDF]

				J. L. Bubenik and D. M. Driscoll Altered RNA Binding Activity Underlies Abnormal Thyroid Hormone Metabolism Linked to a Mutation in Selenocysteine Insertion Sequence-binding Protein 2 J. Biol. Chem., November 30, 2007; 282(48): 34653 - 34662. [Abstract] [Full Text] [PDF]

				K. Caban, S. A. Kinzy, and P. R. Copeland 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., September 15, 2007; 27(18): 6350 - 6360. [Abstract] [Full Text] [PDF]

				M. T. Howard, M. W. Moyle, G. Aggarwal, B. A. Carlson, and C. B. Anderson A recoding element that stimulates decoding of UGA codons by Sec tRNA[Ser]Sec RNA, June 1, 2007; 13(6): 912 - 920. [Abstract] [Full Text] [PDF]

				L. V. Papp, J. Lu, F. Striebel, D. Kennedy, A. Holmgren, and K. K. Khanna The Redox State of SECIS Binding Protein 2 Controls Its Localization and Selenocysteine Incorporation Function. Mol. Cell. Biol., July 1, 2006; 26(13): 4895 - 4910. [Abstract] [Full Text] [PDF]

				L. A. de Jesus, P. R. Hoffmann, T. Michaud, E. P. Forry, A. Small-Howard, R. J. Stillwell, N. Morozova, J. W. Harney, and M. J. Berry Nuclear Assembly of UGA Decoding Complexes on Selenoprotein mRNAs: a Mechanism for Eluding Nonsense-Mediated Decay? Mol. Cell. Biol., March 1, 2006; 26(5): 1795 - 1805. [Abstract] [Full Text] [PDF]

				D. E. Handy, G. Hang, J. Scolaro, N. Metes, N. Razaq, Y. Yang, and J. Loscalzo Aminoglycosides Decrease Glutathione Peroxidase-1 Activity by Interfering with Selenocysteine Incorporation J. Biol. Chem., February 10, 2006; 281(6): 3382 - 3388. [Abstract] [Full Text] [PDF]

				D. E. Handy, Y. Zhang, and J. Loscalzo Homocysteine Down-regulates Cellular Glutathione Peroxidase (GPx1) by Decreasing Translation J. Biol. Chem., April 22, 2005; 280(16): 15518 - 15525. [Abstract] [Full Text] [PDF]