This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Kozerski, C.
Right arrow Articles by Herrmann, A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Kozerski, C.
Right arrow Articles by Herrmann, A.

 Previous Article  |  Next Article 

Journal of Virology, August 2000, p. 7529-7537, Vol. 74, No. 16
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Modification of the Cytoplasmic Domain of Influenza Virus Hemagglutinin Affects Enlargement of the Fusion Pore

Christine Kozerski,1,dagger Evgeni Ponimaskin,2,Dagger Britta Schroth-Diez,1 Michael F. G. Schmidt,2 and Andreas Herrmann1,*

Institut für Biologie/Biophysik, Mathematisch-Naturwissenschaftliche Fakultät I, Humboldt-Universität zu Berlin, D-10115 Berlin,1 and Institut für Immunologie und Molekularbiologie, Fachbereich Veterinärmedizin der Freien Universität Berlin, D-10117 Berlin,2 Germany

Received 7 December 1999/Accepted 4 May 2000

The fusion activity of chimeras of influenza virus hemagglutinin (HA) (from A/fpv/Rostock/34; subtype H7) with the transmembrane domain (TM) and/or cytoplasmic tail (CT) either from the nonviral, nonfusogenic T-cell surface protein CD4 or from the fusogenic Sendai virus F-protein was studied. Wild-type or chimeric HA was expressed in CV-1 cells by the transient T7-RNA-polymerase vaccinia virus expression system. Subsequently, the fusion activity of the expression products was monitored with red blood cells or ghosts as target cells. To assess the different steps of fusion, target cells were labeled with the fluorescent membrane label octadecyl rhodamine B-chloride (R18) (membrane fusion) and with the cytoplasmic fluorophores calcein (molecular weight [MW], 623; formation of small aqueous fusion pore) and tetramethylrhodamine-dextran (MW, 10,000; enlargement of fusion pore). All chimeric HA/F-proteins, as well as the chimera with the TM of CD4 and the CT of HA, were able to mediate the different steps of fusion very similarly to wild-type HA. Quite differently, chimeric proteins with the CT of CD4 were strongly impaired in mediating pore enlargement. However, membrane fusion and formation of small pores were similar to those of wild-type HA, indicating that the conformational change of the ectodomain and earlier fusion steps were not inhibited. Various properties of the CT which may affect pore enlargement are considered. We surmise that the hydrophobicity of the sequence adjacent to the transmembrane domain is important for pore dilation.

* Corresponding author. Mailing address: Humboldt-Universität zu Berlin, Mathematisch-Naturwissenschaftliche Fakultät I, Institut für Biologie/Biophysik, Invalidenstrasse 43, D-10115 Berlin, Germany. Phone: 49-30-2093-8830. Fax: 49-30-2093-8585. E-mail: Andreas=Herrmann{at}rz.hu-berlin.de.

dagger Present address: Department of Cell Biology, University of Virginia School of Medicine, Charlottesville, VA 22908-0732.

Dagger Present address: Zentrum für Physiologie und Pathophysiologie, Abt. Neuro- und Sinnesphysiologie, D-37073 Göttingen, Germany.

Journal of Virology, August 2000, p. 7529-7537, Vol. 74, No. 16
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Barry, C., Duncan, R. (2009). Multifaceted Sequence-Dependent and -Independent Roles for Reovirus FAST Protein Cytoplasmic Tails in Fusion Pore Formation and Syncytiogenesis. J. Virol. 83: 12185-12195 [Abstract] [Full Text]  
  • Ujike, M., Nakajima, K., Nobusawa, E. (2006). A point mutation at the C terminus of the cytoplasmic domain of influenza B virus haemagglutinin inhibits syncytium formation. J. Gen. Virol. 87: 1669-1676 [Abstract] [Full Text]  
  • Li, M., Li, Z.-N., Yao, Q., Yang, C., Steinhauer, D. A., Compans, R. W. (2006). Murine Leukemia Virus R Peptide Inhibits Influenza Virus Hemagglutinin-Induced Membrane Fusion.. J. Virol. 80: 6106-6114 [Abstract] [Full Text]  
  • Nolan, S., Cowan, A. E., Koppel, D. E., Jin, H., Grote, E. (2006). FUS1 Regulates the Opening and Expansion of Fusion Pores between Mating Yeast. Mol. Biol. Cell 17: 2439-2450 [Abstract] [Full Text]  
  • Wagner, R., Herwig, A., Azzouz, N., Klenk, H. D. (2005). Acylation-Mediated Membrane Anchoring of Avian Influenza Virus Hemagglutinin Is Essential for Fusion Pore Formation and Virus Infectivity. J. Virol. 79: 6449-6458 [Abstract] [Full Text]  
  • Zaitseva, E., Mittal, A., Griffin, D. E., Chernomordik, L. V. (2005). Class II fusion protein of alphaviruses drives membrane fusion through the same pathway as class I proteins. JCB 169: 167-177 [Abstract] [Full Text]  
  • Ujike, M., Nakajima, K., Nobusawa, E. (2004). Influence of Acylation Sites of Influenza B Virus Hemagglutinin on Fusion Pore Formation and Dilation. J. Virol. 78: 11536-11543 [Abstract] [Full Text]  
  • Matsuyama, S., Delos, S. E., White, J. M. (2004). Sequential Roles of Receptor Binding and Low pH in Forming Prehairpin and Hairpin Conformations of a Retroviral Envelope Glycoprotein. J. Virol. 78: 8201-8209 [Abstract] [Full Text]  
  • Shmulevitz, M., Salsman, J., Duncan, R. (2003). Palmitoylation, Membrane-Proximal Basic Residues, and Transmembrane Glycine Residues in the Reovirus p10 Protein Are Essential for Syncytium Formation. J. Virol. 77: 9769-9779 [Abstract] [Full Text]  
  • Lin, X., Derdeyn, C. A., Blumenthal, R., West, J., Hunter, E. (2003). Progressive Truncations C Terminal to the Membrane-Spanning Domain of Simian Immunodeficiency Virus Env Reduce Fusogenicity and Increase Concentration Dependence of Env for Fusion. J. Virol. 77: 7067-7077 [Abstract] [Full Text]  
  • Kalia, V., Sarkar, S., Gupta, P., Montelaro, R. C. (2003). Rational Site-Directed Mutations of the LLP-1 and LLP-2 Lentivirus Lytic Peptide Domains in the Intracytoplasmic Tail of Human Immunodeficiency Virus Type 1 gp41 Indicate Common Functions in Cell-Cell Fusion but Distinct Roles in Virion Envelope Incorporation. J. Virol. 77: 3634-3646 [Abstract] [Full Text]  
  • Drummond, S. P., Wilson, K. L. (2002). Interference with the cytoplasmic tail of gp210 disrupts "close apposition" of nuclear membranes and blocks nuclear pore dilation. JCB 158: 53-62 [Abstract] [Full Text]  
  • Sakai, T., Ohuchi, R., Ohuchi, M. (2002). Fatty Acids on the A/USSR/77 Influenza Virus Hemagglutinin Facilitate the Transition from Hemifusion to Fusion Pore Formation. J. Virol. 76: 4603-4611 [Abstract] [Full Text]  
  • Maresova, L., Pasieka, T. J., Grose, C. (2001). Varicella-Zoster Virus gB and gE Coexpression, but Not gB or gE Alone, Leads to Abundant Fusion and Syncytium Formation Equivalent to Those from gH and gL Coexpression. J. Virol. 75: 9483-9492 [Abstract] [Full Text]  
  • Dutch, R. E., Lamb, R. A. (2001). Deletion of the Cytoplasmic Tail of the Fusion Protein of the Paramyxovirus Simian Virus 5 Affects Fusion Pore Enlargement. J. Virol. 75: 5363-5369 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»