TY - JOUR
T1 - N-linked glycosylation of dengue virus NS1 protein modulates secretion, cell-surface expression, hexamer stability, and interactions with human complement
AU - Somnuke, Pawit
AU - Hauhart, Richard E.
AU - Atkinson, John P.
AU - Diamond, Michael S.
AU - Avirutnan, Panisadee
N1 - Funding Information:
We thank W. Klimstra and D. Lenschow for the SINV expression vectors, S. Youn for DENV-2 NS1 expression constructs, D. Fremont, D. Spitzer, E. Miller, C. Nelson, M. Barrow, and S. Youn for experimental help and advice, and C. Puttikhunt and W. Kasinrerk for providing DENV NS1 specific Abs. This work was supported by the Midwest Regional Centers for Excellence for Biodefense and Emerging Infectious Disease Research ( U54-AI057160 to M.S. Diamond and J.P. Atkinson), and the National Center for Genetic Engineering and Biotechnology (BIOTEC), Thailand (P. Avirutnan). P. Somnuke is supported by an M.D.–Ph.D. training fellowship from Medical Scholars Program, Mahidol University. P. Avirutnan has been supported by the National Institutes of Health postdoctoral training grants from the Divisions of Dermatology and Rheumatology in the Department of Medicine, Washington University School of Medicine.
PY - 2011/5/10
Y1 - 2011/5/10
N2 - Dengue virus (DENV) NS1 is a versatile non-structural glycoprotein that is secreted as a hexamer, binds to the cell surface of infected and uninfected cells, and has immune evasive functions. DENV NS1 displays two conserved N-linked glycans at N130 and N207. In this study, we examined the role of these two N-linked glycans on NS1 secretion, stability, and function. Because some groups have reported reduced yields of infectious DENV when N130 and N207 are changed, we analyzed glycosylation-deficient NS1 phenotypes using a transgenic expression system. We show that the N-linked glycan at position 130 is required for stabilization of the secreted hexamer whereas the N-linked glycan at residue 207 facilitates secretion and extracellular protein stability. Moreover, NS1 mutants lacking an N-linked glycan at N130 did not interact efficiently with complement components C1s and C4. In summary, our results elucidate the contribution of N-linked glycosylation to the function of DENV NS1.
AB - Dengue virus (DENV) NS1 is a versatile non-structural glycoprotein that is secreted as a hexamer, binds to the cell surface of infected and uninfected cells, and has immune evasive functions. DENV NS1 displays two conserved N-linked glycans at N130 and N207. In this study, we examined the role of these two N-linked glycans on NS1 secretion, stability, and function. Because some groups have reported reduced yields of infectious DENV when N130 and N207 are changed, we analyzed glycosylation-deficient NS1 phenotypes using a transgenic expression system. We show that the N-linked glycan at position 130 is required for stabilization of the secreted hexamer whereas the N-linked glycan at residue 207 facilitates secretion and extracellular protein stability. Moreover, NS1 mutants lacking an N-linked glycan at N130 did not interact efficiently with complement components C1s and C4. In summary, our results elucidate the contribution of N-linked glycosylation to the function of DENV NS1.
KW - Complement
KW - Dengue virus
KW - Flavivirus
KW - N-linked glycosylation
KW - Non-structural protein nS1
UR - http://www.scopus.com/inward/record.url?scp=79954633991&partnerID=8YFLogxK
U2 - 10.1016/j.virol.2011.02.022
DO - 10.1016/j.virol.2011.02.022
M3 - Article
C2 - 21429549
AN - SCOPUS:79954633991
SN - 0042-6822
VL - 413
SP - 253
EP - 264
JO - Virology
JF - Virology
IS - 2
ER -