Giuseppe
Emeritus
Cell adhesion-dependent membrane trafficking of a binding partner for the ebolavirus glycoprotein is a determinant of viral entry (PNAS, abstract, edited)
[Source: Proc Natl Acad Sci USA, full text: <cite cite="http://www.pnas.org/content/107/38/16637.short?rss=1">Cell adhesion-dependent membrane trafficking of a binding partner for the ebolavirus glycoprotein is a determinant of viral entry ? PNAS</cite>. Abstract, edited.]
Cell adhesion-dependent membrane trafficking of a binding partner for the ebolavirus glycoprotein is a determinant of viral entry
Derek Dube (a), Kathryn L. Schornberg (b), Charles J. Shoemaker (b), Sue E. Delos (b), Tzanko S. Stantchev (c,d), Kathleen A. Clouse (d), Christopher C. Broder (c), and Judith M. White (a,b)
Author Affiliations
Departments of (a) Microbiology and (b) Cell Biology, University of Virginia, Charlottesville, VA 22908;
(c) Department of Microbiology and Immunology, Uniformed Services University, Bethesda, MD 20814; and
(d) Division of Monoclonal Antibodies, Center for Drug Evaluation and Research, US Food and Drug Administration, Bethesda, MD 20892
Edited by Robert A. Lamb, Northwestern University, Evanston, IL, and approved August 10, 2010 (received for review June 15, 2010)
Abstract
Ebolavirus is a hemorrhagic fever virus associated with high mortality. Although much has been learned about the viral lifecycle and pathogenesis, many questions remain about virus entry. We recently showed that binding of the receptor binding region (RBR) of the ebolavirus glycoprotein (GP) and infection by GP pseudovirions increase on cell adhesion independently of mRNA or protein synthesis. One model to explain these observations is that, on cell adhesion, an RBR binding partner translocates from an intracellular vesicle to the cell surface. Here, we provide evidence for this model by showing that suspension 293F cells contain an RBR binding site within a membrane-bound compartment associated with the trans-Golgi network and microtubule-organizing center. Consistently, trafficking of the RBR binding partner to the cell surface depends on microtubules, and the RBR binding partner is internalized when adherent cells are placed in suspension. Based on these observations, we reexamined the claim that lymphocytes, which are critical for ebolavirus pathogenesis, are refractory to infection because they lack an RBR binding partner. We found that both cultured and primary human lymphocytes (in suspension) contain an intracellular pool of an RBR binding partner. Moreover, we identified two adherent primate lymphocytic cell lines that bind RBR at their surface and strikingly, support GP-mediated entry and infection. In summary, our results reveal a mode of determining viral entry by a membrane-trafficking event that translocates an RBR binding partner to the cell surface, and they suggest that this process may be operative in cells important for ebolavirus pathogenesis (e.g., lymphocytes and macrophages).
* filovirus
* negative strand RNA virus
* virus receptor
* lymphocytes
* macrophages
Footnotes
To whom correspondence should be addressed. E-mail: jw7g@virginia.edu.
Author contributions: D.D., K.L.S., S.E.D., C.C.B., and J.M.W. designed research; D.D., K.L.S., C.J.S., and T.S.S. performed research; C.J.S., T.S.S., and K.A.C. contributed new reagents/analytic tools; D.D., K.L.S., and J.M.W. analyzed data; and D.D., K.L.S., S.E.D., T.S.S., K.A.C., C.C.B., and J.M.W. wrote the paper.
The authors declare no conflict of interest.
This article is a PNAS Direct Submission.
This article contains supporting information online at www.pnas.org/lookup/suppl/doi:10.1073/pnas.1008509107/-/DCSupplemental.
Freely available online through the PNAS open access option.
-
-----<cite cite="http://www.pnas.org/content/107/38/16637.short?rss=1"></cite>
[Source: Proc Natl Acad Sci USA, full text: <cite cite="http://www.pnas.org/content/107/38/16637.short?rss=1">Cell adhesion-dependent membrane trafficking of a binding partner for the ebolavirus glycoprotein is a determinant of viral entry ? PNAS</cite>. Abstract, edited.]
Cell adhesion-dependent membrane trafficking of a binding partner for the ebolavirus glycoprotein is a determinant of viral entry
Derek Dube (a), Kathryn L. Schornberg (b), Charles J. Shoemaker (b), Sue E. Delos (b), Tzanko S. Stantchev (c,d), Kathleen A. Clouse (d), Christopher C. Broder (c), and Judith M. White (a,b)
Author Affiliations
Departments of (a) Microbiology and (b) Cell Biology, University of Virginia, Charlottesville, VA 22908;
(c) Department of Microbiology and Immunology, Uniformed Services University, Bethesda, MD 20814; and
(d) Division of Monoclonal Antibodies, Center for Drug Evaluation and Research, US Food and Drug Administration, Bethesda, MD 20892
Edited by Robert A. Lamb, Northwestern University, Evanston, IL, and approved August 10, 2010 (received for review June 15, 2010)
Abstract
Ebolavirus is a hemorrhagic fever virus associated with high mortality. Although much has been learned about the viral lifecycle and pathogenesis, many questions remain about virus entry. We recently showed that binding of the receptor binding region (RBR) of the ebolavirus glycoprotein (GP) and infection by GP pseudovirions increase on cell adhesion independently of mRNA or protein synthesis. One model to explain these observations is that, on cell adhesion, an RBR binding partner translocates from an intracellular vesicle to the cell surface. Here, we provide evidence for this model by showing that suspension 293F cells contain an RBR binding site within a membrane-bound compartment associated with the trans-Golgi network and microtubule-organizing center. Consistently, trafficking of the RBR binding partner to the cell surface depends on microtubules, and the RBR binding partner is internalized when adherent cells are placed in suspension. Based on these observations, we reexamined the claim that lymphocytes, which are critical for ebolavirus pathogenesis, are refractory to infection because they lack an RBR binding partner. We found that both cultured and primary human lymphocytes (in suspension) contain an intracellular pool of an RBR binding partner. Moreover, we identified two adherent primate lymphocytic cell lines that bind RBR at their surface and strikingly, support GP-mediated entry and infection. In summary, our results reveal a mode of determining viral entry by a membrane-trafficking event that translocates an RBR binding partner to the cell surface, and they suggest that this process may be operative in cells important for ebolavirus pathogenesis (e.g., lymphocytes and macrophages).
* filovirus
* negative strand RNA virus
* virus receptor
* lymphocytes
* macrophages
Footnotes
To whom correspondence should be addressed. E-mail: jw7g@virginia.edu.
Author contributions: D.D., K.L.S., S.E.D., C.C.B., and J.M.W. designed research; D.D., K.L.S., C.J.S., and T.S.S. performed research; C.J.S., T.S.S., and K.A.C. contributed new reagents/analytic tools; D.D., K.L.S., and J.M.W. analyzed data; and D.D., K.L.S., S.E.D., T.S.S., K.A.C., C.C.B., and J.M.W. wrote the paper.
The authors declare no conflict of interest.
This article is a PNAS Direct Submission.
This article contains supporting information online at www.pnas.org/lookup/suppl/doi:10.1073/pnas.1008509107/-/DCSupplemental.
Freely available online through the PNAS open access option.
-
-----<cite cite="http://www.pnas.org/content/107/38/16637.short?rss=1"></cite>