Dengue virus-induced modifications of host cell membranes

Abstract

Enzymatic markers and electron microscopy were utilized to determine the cellular origin of the membrane types isolated from type 2 dengue virus-infected BHK cells by discontinouous sucrose gradient centrifugation. The results showed an apparent separation of plasma membrane, smooth and rough endoplasmic reticulum with increasing density. Virus-induced protein and RNA synthesis, as indicated by the incorporation of radiolabeled precursors, was localized on the rough endoplasmic reticulum. Glycosylation, measured by the incorporation of radiolabeled glucosamine into membrane-associated proteins, was most active in the bands of intermediate and smooth endoplasmic reticulum. Polyacrylamide gel electrophoresis of isolated membrane bands, radiolabeled in the presence of actinomycin D, after pulse inhibition by cycloheximide, revealed seven virusspecified proteins associated with all membrane fractions. Viral structural protein V-3, and nonstructural proteins NV-3 and NV-2, increased with decreasing density, whereas NV-5 and NV-4 remained constant. The viral capsid protein V-2 was depleted in the intermediate and smooth endoplasmic reticulum, suggesting that these membranes may serve as the sites for viral maturation. NV-3 was the most prominent virus-specified protein found in the plasma membrane.

Morphogenesis of Dengue‐1 Virus in Cultures of a Human Leukemic Leukocyte Line (J‐111)

“…Some of these images are essentially the same as those reported by previous investigators dealing with various types of cultured cells infected with dengue viruses, e.g., Vero and BHK (6,7,10,14), LLCMK2 (2)(3)(4)9), Raji and related human lymphoblastoid cells (15,16), as well as infected mouse brain cells (12,13). Regarded as particularly significant was observation of "budding" particles.…”

Structural disorder in the proteome and interactome of Alkhurma virus (ALKV)

“…As it was already indicated, similar to other flaviviral proteins, all ALKV proteins (three structural proteins, envelope, E; membrane precursor, PrM; capsid C, and seven non-structural (NS) proteins, NS1, NS2A, NS2B, NS3, NS4A, NS4B/2k, and NS5) are generated as a result of the co-and post-translational cleavage of the genomic polyprotein encoded by a single ORF. The viral genome is translated into the viral polyprotein on the rough endoplasmic reticulum (ER) [92]. NS1, prM, and E are translated into the ER lumen, the transmembrane domains of NS2A, NS2B, NS4A, and NS4B are translated into the ER membrane, and proteins C, NS3, and NS5 remain in the cytoplasm [12,93].…”

Ultrastructure of the replication sites of positive-strand RNA viruses

“…Molecular aspects of DENV replication are elaborated in other reviews (Bartenschlager and Miller, 2008;Acosta et al, 2014). First visualizations of membrane rearrangements within DENV-infected cells showed cytoplasmic vacuoles and accumulation of one or several virions in crystalloid arrays, which are surrounded by host membranes (Stohlman et al, 1975;Matsumura et al, 1971). Later studies focused more on infected mosquito cells and visualized different steps of the viral life cycle (Hase et al, 1987;Tu et al, 1998).…”