Fluorescently Labeled Human Papillomavirus Pseudovirions for Use in Virus Entry Experiments

Current Protocols in Microbiology
Pilar Samperio Ventayol, Mario Schelhaas

Abstract

Human papillomaviruses (HPV) infect skin or mucosal epidermis. The simplistic capsid consists of a major capsid protein L1, a minor capsid protein L2, and a double-stranded circular DNA of about 8 kB in size. The development of HPV-based vectors [i.e., pseudovirions (PsV)] as tools to study the initial infection has facilitated our understanding of HPV entry. The covalent coupling of fluorescent molecules to these PsV allows following the viruses en route to the nucleus, i.e., the site of replication. In the first section, we describe a facile method to covalently label HPV PsV that retain their infectivity. In this method, fluorophores coupled to a reactive succinimidyl ester are covalently attached to amine residues in the virion in a one-step chemical reaction. In the second section of this unit, several assays are outlined that use the fluorescently labeled virions for entry studies in live and fixed cells.

References

Feb 1, 1980·The Journal of Cell Biology·A HeleniusE Fries
Jun 4, 1993·Journal of Immunological Methods·C P WanB H Lau
Jan 11, 2001·Journal of Virology·T GiroglouM Sapp
Feb 18, 2004·Virology·Timothy D Culp, Neil D Christensen
Aug 6, 2005·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Robert M MartinM Cristina Cardoso
Jan 11, 2007·Journal of Microscopy·S Bolte, F P Cordelières
Jan 30, 2008·Current Protocols in Cell Biology·Christopher B Buck, Cynthia D Thompson
Feb 11, 2009·Cell·Patrick SteigemannDaniel W Gerlich
Nov 3, 2009·The FEBS Journal·Martin Sapp, Malgorzata Bienkowska-Haba
Feb 25, 2011·Journal of Virology·Sabrina EngelAri Helenius
Feb 22, 2012·Methods in Enzymology·Helge Ewers, Mario Schelhaas
Apr 10, 2013·Proceedings of the National Academy of Sciences of the United States of America·Alex LipovskyDaniel DiMaio
Apr 26, 2013·Journal of Virology·Gilles SpodenMario Schelhaas
Feb 15, 2014·Current Opinion in Virology·Patricia M Day, Mario Schelhaas

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.