Determinants for membrane association and permeabilization of the coxsackievirus 2B protein and the identification of the Golgi complex as the target organelle.
The 2B protein of enterovirus is responsible for the alterations in the permeability of secretory membranes and the plasma membrane in infected cells. The structural requirements for the membrane association and the subcellular localization of this essential virus protein, however, have not been defined. Here, we provide evidence that the 2B protein is an integral membrane protein in vivo that is predominantly localized at the Golgi complex upon individual expression. Addition of organelle-specific targeting signals to the 2B protein revealed that the Golgi localization is an absolute prerequisite for the ability of the protein to modify plasma membrane permeability. Expression of deletion mutants and heterologous proteins containing specific domains of the 2B protein demonstrated that each of the two hydrophobic regions could mediate membrane binding individually. However, the presence of both hydrophobic regions was required for the correct membrane association, efficient Golgi targeting, and the membrane-permeabilizing activity of the 2B protein, suggesting that the two hydrophobic regions are cooperatively involved in the formation of a membrane-integral complex. The formation of membrane-integral pores by the 2B protein in the Golgi complex and the possible mechanism by which a Golgi-localized virus protein modifies plasma membrane permeability are discussed.
de Jong
Arjan S
AS
Department of Medical Microbiology, Nijmegen Center for Molecular Life Sciences, University Medical Center Nijmegen, P. O. Box 9100, The Netherlands.
Wessels
Els
E
Dijkman
Henri B P M
HB
Galama
Jochem M D
JM
Melchers
Willem J G
WJ
Willems
Peter H G M
PH
van Kuppeveld
Frank J M
FJ
D000595:Amino Acid Sequence; D000818:Animals; D002462:Cell Membrane / Q000737:chemistry*; D002463:Cell Membrane Permeability; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D006056:Golgi Apparatus / Q000737:chemistry*; D008969:Molecular Sequence Data; D014764:Viral Proteins / Q000032:analysis / Q000737:chemistry* / Q000502:physiology; D014779:Virus Replication
D014764:Viral Proteins; C097017:coxsackie B3 virus protein 2B