Fragmentation of the Golgi apparatus provides replication membranes for human rhinovirus 1A

Abstract

All viruses with a positive-stranded RNA genome replicate their genomic RNA in association with membranes from the host cell. Here we demonstrate a novel organelle source of replication membranes for human rhinovirus 1A (HRV-1A). HRV-1A infection induces fragmentation of the Golgi apparatus, and Golgi membranes are rearranged into vesicles of approximately 250-500 nm diameter. The newly distributed Golgi membranes co-localize with viral RNA replication templates, strongly suggesting that the observed vesicles are the sites of viral RNA replication. Expression of the HRV-1A 3A protein induces alterations in the Golgi staining pattern similar to those seen during viral infection, and expressed 3A localizes to the Golgi-derived membranes. Taken together, these data show that in HRV-1A infection, the 3A protein plays a role in fragmenting the Golgi complex and generating vesicles that are used as the site of viral RNA replication.

Human Rhinovirus 2 Induces the Autophagic Pathway and Replicates More Efficiently in Autophagic Cells

“…Rapamycin induction caused approximately 70% of cells in the population to display visible puncta, while PV and HRV-2 induced about half of the cells to display visible puncta. These numbers are in line with previous data for PV, indicating that HRV-2 and PV score equivalently in this assay (23). Therefore, by assaying formation of punctate LC3 structures, we observed that HRV-2 appears to stimulate autophagosome formation.…”

“…PV infection or rapamycin treatment induced formation of puncta; arrows indicate a representative cell in each field with visible puncta. HRV-1A infection did not induce formation of puncta, in agreement with our previous work, indicating that formation of puncta is not a general response to picornavirus infection (23). Infection by HRV-2, however, induces formation of puncta similar to that seen during PV infection.…”

“…Also as expected, the yield of HRV-1A was unaffected by drug treatment (23). We found that the HRV-2 yield paralleled that of PV closely, increasing 2-to 3-fold with rapamycin treatment and decreasing by Ͼ1 log with 3-MA treatment.…”

“…We transiently transfected 293T cells with an enhanced GFP (EGFP)-LC3-expressing vector as described previously and, 24 h later, infected cells for 6 h with PV, HRV-1A, or HRV-2 at a multiplicity of infection (MOI) of 50 (23). For controls, we mock infected cells or treated cells for 6 h with either 10 M rapamycin or an equivalent volume of dimethyl sulfoxide (DMSO) carrier.…”

Role of Phosphatidylinositol 4-Phosphate (PI4P) and Its Binding Protein GOLPH3 in Hepatitis C Virus Secretion

“…The fragmentation of the Golgi in HCV correlates with the decrease in general secretory capacity of the infected cell but further implicates the organelle in important viral processes and hints toward sequestration of secretory components at sites of viral assembly. Golgi disruption and decrease in secretory capacity of host cells is a peculiar feature of many infectious agents such as Chlamydia, poliovirus, and rhinovirus 1A, where the Golgi membranes serve an important role in viral replication and morphogenesis (57)(58)(59)(60).…”