Pathways such as macropinocytosis which is the canonical pathway for EBOV entry. Our data indicate that MBL also mediates internalization of virus via macropinocytosis but suggests that MBL-mediated uptake preferentially utilizes microtubules compared with the canonical EBOV pathway which is dependent on both microtubules and actin. doi:10.1371/journal.pone.0060838.gcirculating C4 [88], we specula
Uction assay). Shown are average adjusted luciferase results from two experiments each with four replicates. The maximal reduction of viral infection was 17fold in the presence of rhMBL, whereas there was no inhibition of infection in the absence of MBL. (EPS)Figure S5 MBL-mediated macropinocytosis of HIVEBOV GP may bypass early endosomes and is dynamin independent. We preincubated HEK293F cells w
Particles mixed withFigure 8. Proposed model of MBL-mediated macropinocytosis of EBOV. MBL carbohydrate recognition domains (CRD) bind to highly glycosylated mucin-rich regions of EBOV GP and the MBL-virion complex is presented to the cell surface. Then MBL binds to cognate cellular receptors, such as C1QBP or calreticulin [6] via MBL collagenous stalks. In this manner, MBL concentrates virus at t
Particles mixed withFigure 8. Proposed model of MBL-mediated macropinocytosis of EBOV. MBL carbohydrate recognition domains (CRD) bind to highly glycosylated mucin-rich regions of EBOV GP and the MBL-virion complex is presented to the cell surface. Then MBL binds to cognate cellular receptors, such as C1QBP or calreticulin [6] via MBL collagenous stalks. In this manner, MBL concentrates virus at t
Ge activity .500 U/mL (13/21 vs 1/ 14, p,0.005) compared with MBL2 O/O or O/A haplotypes (O refers to B,C or D alleles). (EPS) Figure S3 Endoglycosidases cleave N-linked glycans in HIV-EBOV GP. We preincubated HIV-EBOZ GP virion-like particles (12,000 pg/ml) with PNGase F or endo H (10,000 U/ml each) diluted in DMEM or with DMEM alone for 1 hour at 37uC. Viruses then underwent gel electrophoresis;
Ge activity .500 U/mL (13/21 vs 1/ 14, p,0.005) compared with MBL2 O/O or O/A haplotypes (O refers to B,C or D alleles). (EPS) Figure S3 Endoglycosidases cleave N-linked glycans in HIV-EBOV GP. We preincubated HIV-EBOZ GP virion-like particles (12,000 pg/ml) with PNGase F or endo H (10,000 U/ml each) diluted in DMEM or with DMEM alone for 1 hour at 37uC. Viruses then underwent gel electrophoresis;
On of fetal calf or fetal bovine serum typically used for EBOV infectivity assays [17,18,44,45,67]. Given that C4 gene copy number variations may lead to a proportionate reduction ofLectin-Dependent Enhancement of Ebola Virusesiological doses of MBL products or blood products with high MBL concentrations to individuals in the setting of infectious diseases and relative hypocomplementemia may be de
Exposed to autoradiography film. High-mannose moieties were preserved only in untreated virus confirming that PNGase F and endo H effectively cleaved high-mannose residues on GPs of HIV-EBOV virion-like particles. (EPS) Figure S4 Thermolysin treatment of HIV-EBOV GP abrogates enhancement of infection by rhMBL in a thermolysin-concentration dependent manner. We preincubated HIV-EBOV GP virion-like