Related Experiment Video
Updated: Jun 20, 2026
11:40
Simplified Reverse Genetics Method to Recover Recombinant Rotaviruses Expressing Reporter Proteins
Published on: April 17, 2020
VP5* rearranges when rotavirus uncoats
Joshua D Yoder1, Shane D Trask, T Phuoc Vo
1Laboratory of Molecular Medicine, Children's Hospital, Boston, Massachusetts 02115, USA.
Journal of Virology
|August 21, 2009
Summary
Rotavirus entry involves a conformational change in its VP4 spike protein. This rearrangement of VP5* occurs during uncoating, facilitating viral infection and membrane penetration.
Area of Science:
- Virology
- Structural Biology
- Molecular Biology
Background:
- Rotavirus requires trypsin cleavage of its VP4 spike protein into VP8* and VP5* for efficient infectivity.
- A recombinant VP5* fragment exhibits a folded-back trimeric structure, suggesting a potential rearrangement mechanism for membrane penetration during viral entry.
Purpose of the Study:
- To investigate the conformational changes of the VP4 spike protein during rotavirus entry.
- To detect the inferred rearrangement of virion-associated VP5* using conformation-specific antibodies.
Main Methods:
- Generation of conformation-specific monoclonal antibodies against a recombinant VP5* fragment in its putative post-membrane penetration conformation.
- Utilizing these antibodies to detect conformational changes in rotavirus VP5* during uncoating.
Main Results:
- A conformational change in the cleaved VP4 spike protein was detected during rotavirus uncoating.
- This conformational change yields a rearranged VP5* structure, confirming the inferred rearrangement.
Conclusions:
- Rotavirus uncoating triggers a significant conformational alteration in the VP4 spike protein.
- This rearrangement of VP5* is crucial for mediating rotavirus membrane penetration and infectivity.
Related Concept Videos
00:57Viral 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.
01:33Coat Assembly and GTPases
Vesicles incorporate different coat protein subunits in different cell locations, which changes the properties of the coat, such as the shape and geometry of the transport vesicles. Thus, vesicle coat proteins also play a significant role in cargo selection.
Coat assembly depends on the local availability of phosphatidylinositol phosphates or PIPs and GTP-binding proteins. Adaptor proteins, which link the coat proteins to the membrane, bind to these PIPs and play a crucial role in controlling...
Coat assembly depends on the local availability of phosphatidylinositol phosphates or PIPs and GTP-binding proteins. Adaptor proteins, which link the coat proteins to the membrane, bind to these PIPs and play a crucial role in controlling...
01:10Retrovirus 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...
01:19Inhibitors 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...
01:26Size and Structure of Viral Genomes
Viral genomes exhibit remarkable diversity in size, structure, and composition, influencing their replication strategies and interactions with host cells. These genomes consist of either DNA or RNA and may be linear or circular. Additionally, they can be single-stranded or double-stranded, with each configuration affecting how the virus propagates within a host. RNA viruses, for instance, generally have smaller genomes than DNA viruses, a factor that contributes to their high mutation rates and...
00:36Viral Mutations
A mutation is a change in the sequence of bases of DNA or RNA in a genome. Some mutations occur during replication of the genome due to errors made by the polymerase enzymes that replicate DNA or RNA. Unlike DNA polymerase, RNA polymerase is prone to errors because it is not capable of “proofreading” its work. Viruses with RNA-based genomes, like HIV, therefore accrue mutations faster than viruses with DNA-based genomes. Because mutation and recombination provide the raw material for adaptive...