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Related Concept Videos

Retrovirus Life Cycles01:10

Retrovirus Life Cycles

Retroviruses have a single-stranded RNA genome that undergoes a special form of replication. Once the retrovirus has entered the host cell, an enzyme called reverse transcriptase synthesizes double-stranded DNA from the retroviral RNA genome. This DNA copy of the genome is then integrated into the host’s genome inside the nucleus via an enzyme called integrase. Consequently, the retroviral genome is transcribed into RNA whenever the host’s genome is transcribed, allowing the retrovirus to...
Retroviruses02:33

Retroviruses

Retroviruses and retrotransposons both insert copies of their genetic elements into the genome of the host cell. Thus, the viral genes are passed on when the host genome is replicated or translated. A typical retroviral DNA sequence contains 3-4 genes that encode the different proteins required for its structural assembly and function as a molecular parasite. This DNA is transcribed into a single mRNA, which is very similar in structure to conventional mRNAs, i.e., it is capped at the 5’...
Inhibitors of Virion Maturation and Assembly01:19

Inhibitors of Virion Maturation and Assembly

As part of their replication cycle, certain viruses synthesize long precursor proteins called polyproteins within infected host cells. In human immunodeficiency virus (HIV), two major polyproteins are produced: Gag and Gag-Pol. The Gag polyprotein supplies the structural components of the virus, while Gag-Pol includes essential viral enzymes such as reverse transcriptase, integrase, and protease. After synthesis, these polyproteins move to the host cell membrane, where they assemble into an...
Subviral Agents01:29

Subviral Agents

Subviral agents are infectious entities that resemble viruses but lack one or more viral components, such as a capsid or essential replication machinery. These agents include viroids, prions, and satellites, each possessing distinct structural and functional characteristics that influence their mode of infection and replication.Viroids are the simplest subviral agents, consisting of circular, single-stranded RNA molecules without a protein coat. They exclusively infect plants, relying entirely...
Viral Recombination00:57

Viral Recombination

Cells are sometimes infected by more than one virus at once. When two viruses disassemble to expose their genomes for replication in the same cell, similar regions of their genomes can pair together and exchange sequences in a process called recombination. Alternatively, viruses with segmented genomes can swap segments in a process called reassortment.
Protein Complex Assembly02:41

Protein Complex Assembly

Proteins can form homomeric complexes with another unit of the same protein or heteromeric complexes with different types.  Most protein complexes self-assemble spontaneously via ordered pathways, while some proteins need assembly factors that guide their proper assembly. Despite the crowded intracellular environment, proteins usually interact with their correct partners and form functional complexes.
Many viruses self-assemble into a fully functional unit using the infected host cell to...

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Development of a Hepatitis B Virus Reporter System to Monitor the Early Stages of the Replication Cycle
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Functional Sindbis virus replicative complexes are formed at the plasma membrane.

Elena I Frolova1, Rodion Gorchakov, Larisa Pereboeva

  • 1Department of Microbiology, University of Alabama, Birmingham, AL 35294-2170, USA. ivfrolov@UAB.edu

Journal of Virology
|September 10, 2010
PubMed
Summary

Sindbis virus replicative complexes form at the plasma membrane in vertebrate cells, not in endosomes as previously thought. This discovery impacts understanding of alphavirus replication and pathogenesis.

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Isolation of Viral Replication Compartment-enriched Sub-nuclear Fractions from Adenovirus-infected Normal Human Cells

Published on: November 12, 2015

Area of Science:

  • Virology
  • Molecular Biology
  • Cell Biology

Background:

  • Alphavirus replication involves forming replicative complexes (RCs).
  • Previously, type 1 cytopathic vacuoles (CPVs) were proposed as sites for RNA synthesis.
  • CPVs feature membrane invaginations (spherules) and accumulate viral nonstructural proteins (nsPs).

Purpose of the Study:

  • To investigate the precise location and formation mechanism of Sindbis virus (SINV) replicative complexes.
  • To challenge the established model of RCs forming within endosomes.

Main Methods:

  • Immunofluorescence microscopy to observe nsP and dsRNA localization.
  • Analysis of viral RNA synthesis in the presence of endocytosis inhibitors.
  • Comparative studies in vertebrate and mosquito cells.

Main Results:

  • SINV RCs, containing dsRNA and nsPs, initially form at the plasma membrane in vertebrate cells.
  • dsRNA intermediates are packaged into membrane spherules on the external plasma membrane surface.
  • RC formation requires nsP123 and nsP4, with dsRNA synthesis being a prerequisite.
  • Plasma membrane-associated RCs are active sites of ssRNA synthesis.
  • RCs primarily remain at the plasma membrane, with minimal relocalization to endosomes late in infection.
  • Endocytosis inhibition has little effect on SINV RNA replication.
  • Mosquito cells show RC association with both plasma and endosomal membranes.

Conclusions:

  • SINV RCs initiate at the plasma membrane in vertebrate cells, contrary to previous hypotheses.
  • Plasma membrane spherules serve as the primary sites for SINV RNA synthesis.
  • Mechanisms of RC formation may differ between insect and vertebrate cells.