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

J. Biol. Chem., Vol. 279, Issue 19, 19924-19935, May 7, 2004

This Article Full Text Full Text (PDF) All Versions of this Article: 279/19/19924    most recent M314094200v1 Submit a Letter to Editor Alert me when this article is cited Alert me when eLetters are posted 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 Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by de Jong, A. S. Articles by van Kuppeveld, F. J. M. Search for Related Content PubMed PubMed Citation Articles by de Jong, A. S. Articles by van Kuppeveld, F. J. M. Social Bookmarking                      What's this?

Mutational Analysis of Different Regions in the Coxsackievirus 2B Protein

REQUIREMENTS FOR HOMO-MULTIMERIZATION, MEMBRANE PERMEABILIZATION, SUBCELLULAR LOCALIZATION, AND VIRUS REPLICATION^*

Arjan S. de Jong, Willem J. G. Melchers, Dirk H. R. F. Glaudemans, Peter H. G. M. Willems, and Frank J. M. van Kuppeveld¶

From the Departments of Medical Microbiology and Biochemistry, Nijmegen Center for Molecular Life Sciences, University Medical Center Nijmegen, 6500 HB Nijmegen, The Netherlands

The coxsackievirus 2B protein is a small hydrophobic protein (99 amino acids) that increases host cell membrane permeability, possibly by forming homo-multimers that build membrane-integral pores. Previously, we defined the functional role of the two hydrophobic regions HR1 and HR2. Here, we investigated the importance of regions outside HR1 and HR2 for multimerization, increasing membrane permeability, subcellular localization, and virus replication through analysis of linker insertion and substitution mutants. From these studies, the following conclusions could be drawn. (i) The hydrophilic region (⁵⁸RNHDD⁶²) between HR1 and HR2 is critical for multimerization and increasing membrane permeability. Substitution analysis of Asn⁶¹ and Asn⁶² demonstrated the preference for short polar side chains (Asp, Asn), residues that are often present in turns, over long polar side chains (Glu, Gln). This finding supports the idea that the hydrophilic region is involved in pore formation by facilitating a turn between HR1 and HR2 to reverse chain direction. (ii) Studies undertaken to define the downstream boundary of HR2 demonstrated that the aromatic residues Trp⁸⁰ and Trp⁸², but not the positively charged residues Arg⁸¹, Lys⁸⁴, and Lys⁸⁶ are important for increasing membrane permeability. (iii) The N terminus is not required for multimerization but does contribute to the membraneactive character of 2B. (iv) The subcellular localization of 2B does not rely on regions outside HR1 and HR2 and does not require multimerization. (v) Virus replication requires both the membrane-active character and an additional function of 2B that is not connected to this activity.

Received for publication, December 23, 2003 , and in revised form, February 18, 2004.

^* This work was partly supported by Netherlands Organization for Scientific Research Grant NWO-917.46.305 and European Communities Grant INTAS 2012. 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.

^¶ To whom correspondence should be addressed: Dept. of Medical Microbiology, Nijmegen Center for Molecular Life Sciences, University Medical Center Nijmegen, P.O. Box 9101, 6500 HB Nijmegen, The Netherlands. Tel.: 3124-361-7574; Fax: 3124-361-4666; E-mail: f.vankuppeveld{at}ncmls.kun.nl.

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

This article has been cited by other articles:

				A. S. de Jong, F. de Mattia, M. M. Van Dommelen, K. Lanke, W. J. G. Melchers, P. H. G. M. Willems, and F. J. M. van Kuppeveld Functional Analysis of Picornavirus 2B Proteins: Effects on Calcium Homeostasis and Intracellular Protein Trafficking J. Virol., April 1, 2008; 82(7): 3782 - 3790. [Abstract] [Full Text] [PDF]

				C. T. Cornell, W. B. Kiosses, S. Harkins, and J. L. Whitton Coxsackievirus B3 Proteins Directionally Complement Each Other To Downregulate Surface Major Histocompatibility Complex Class I J. Virol., July 1, 2007; 81(13): 6785 - 6797. [Abstract] [Full Text] [PDF]

				D. Clarke, S. Griffin, L. Beales, C. St. Gelais, S. Burgess, M. Harris, and D. Rowlands Evidence for the Formation of a Heptameric Ion Channel Complex by the Hepatitis C Virus P7 Protein in Vitro J. Biol. Chem., December 1, 2006; 281(48): 37057 - 37068. [Abstract] [Full Text] [PDF]

				A. S. de Jong, H.-J. Visch, F. de Mattia, M. M. van Dommelen, H. G. Swarts, T. Luyten, G. Callewaert, W. J. Melchers, P. H. Willems, and F. J. van Kuppeveld The Coxsackievirus 2B Protein Increases Efflux of Ions from the Endoplasmic Reticulum and Golgi, thereby Inhibiting Protein Trafficking through the Golgi J. Biol. Chem., May 19, 2006; 281(20): 14144 - 14150. [Abstract] [Full Text] [PDF]

				C.-K. Lee, K. Kono, E. Haas, K.-S. Kim, K. M. Drescher, N. M. Chapman, and S. Tracy Characterization of an infectious cDNA copy of the genome of a naturally occurring, avirulent coxsackievirus B3 clinical isolate J. Gen. Virol., January 1, 2005; 86(1): 197 - 210. [Abstract] [Full Text] [PDF]