Principles of polyoma- and papillomavirus uncoating

Cerqueira, Carla; Schelhaas, Mario

doi:10.1007/s00430-012-0262-1

Cited by 17 publications

(11 citation statements)

References 131 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Uncoating is an important step in HPV entry that releases the L2/vDNA complex from the L1 capsid to allow for subsequent trafficking to the TGN and the nucleus [43]. Breakdown of the HPV capsid requires endosome acidification [15, 19, 20] and dissociation of L2/vDNA from L1 pentamers requires the activity of host cyclophilins [22].…”

Section: Resultsmentioning

confidence: 99%

Translocation of the papillomavirus L2/vDNA complex across the limiting membrane requires the onset of mitosis

Calton¹,

Bronnimann²,

Manson³

et al. 2017

PLoS Pathog

114

View full text Add to dashboard Cite

The human papillomavirus type 16 (HPV16) L2 protein acts as a chaperone to ensure that the viral genome (vDNA) traffics from endosomes to the trans-Golgi network (TGN) and eventually the nucleus, where HPV replication occurs. En route to the nucleus, the L2/vDNA complex must translocate across limiting intracellular membranes. The details of this critical process remain poorly characterized. We have developed a system based on subcellular compartmentalization of the enzyme BirA and its cognate substrate to detect membrane translocation of L2-BirA from incoming virions. We find that L2 translocation requires transport to the TGN and is strictly dependent on entry into mitosis, coinciding with mitotic entry in synchronized cells. Cell cycle arrest causes retention of L2/vDNA at the TGN; only release and progression past G2/M enables translocation across the limiting membrane and subsequent infection. Microscopy of EdU-labeled vDNA reveals a rapid and dramatic shift in vDNA localization during early mitosis. At late G2/early prophase vDNA egresses from the TGN to a pericentriolar location, accumulating there through prometaphase where it begins to associate with condensed chromosomes. By metaphase and throughout anaphase the vDNA is seen bound to the mitotic chromosomes, ensuring distribution into both daughter nuclei. Mutations in a newly defined chromatin binding region of L2 potently blocked translocation, suggesting that translocation is dependent on chromatin binding during prometaphase. This represents the first time a virus has been shown to functionally couple the penetration of limiting membranes to cellular mitosis, explaining in part the tropism of HPV for mitotic basal keratinocytes.

show abstract

Section: Resultsmentioning

confidence: 99%

Translocation of the papillomavirus L2/vDNA complex across the limiting membrane requires the onset of mitosis

Calton¹,

Bronnimann²,

Manson³

et al. 2017

PLoS Pathog

114

View full text Add to dashboard Cite

show abstract

“…Generally, uncoating occurs through interaction with the cellular environment, i.e., different chemical milieus or specific interactions with cellular proteins. Thereby, stabilizing contacts in the virus structure are modified through conformational changes, isomerization of covalent bonds, refolding, and/or proteolysis (11).…”

mentioning

confidence: 99%

Kallikrein-8 Proteolytically Processes Human Papillomaviruses in the Extracellular Space To Facilitate Entry into Host Cells

Cerqueira

Ventayol

Vogeley

et al. 2015

J Virol

Self Cite

104

View full text Add to dashboard Cite

The entry of human papillomaviruses into host cells is a complex process. It involves conformational changes at the cell surface, receptor switching, internalization by a novel endocytic mechanism, uncoating in endosomes, trafficking of a subviral complex to the Golgi complex, and nuclear entry during mitosis. Here, we addressed how the stabilizing contacts in the capsid of human papillomavirus 16 (HPV16) may be reversed to allow uncoating of the viral genome. Using biochemical and cell-biological analyses, we determined that the major capsid protein L1 underwent proteolytic cleavage during entry. In addition to a dispensable cathepsin-mediated proteolysis that occurred likely after removal of capsomers from the subviral complex in endosomes, at least two further proteolytic cleavages of L1 were observed, one of which was independent of the low-pH environment of endosomes. This cleavage occurred extracellularly. Further analysis showed that the responsible protease was the secreted trypsin-like serine protease kallikrein-8 (KLK8) involved in epidermal homeostasis and wound healing. Required for infection, the cleavage was facilitated by prior interaction of viral particles with heparan sulfate proteoglycans. KLK8-mediated cleavage was crucial for further conformational changes exposing an important epitope of the minor capsid protein L2. Occurring independently of cyclophilins and of furin that mediate L2 exposure, KLK8-mediated cleavage of L1 likely facilitated access to L2, located in the capsid lumen, and potentially uncoating. Since HPV6 and HPV18 also required KLK8 for entry, we propose that the KLK8-dependent entry step is conserved. IMPORTANCEOur analysis of the proteolytic processing of incoming HPV16, an etiological agent of cervical cancer, demonstrated that the capsid is cleaved extracellularly by a serine protease active during wound healing and that this cleavage was crucial for infection. The cleavage of L1 is one of at least four structural alterations that prime the virus extracellularly for receptor switching, internalization, and possibly uncoating. This step was also important for HPV6 and HPV18, which may suggest that it is conserved among the papillomaviruses. This study advances the understanding of how HPV16 initially infects cells, strengthens the notion that wounding facilitates infection of epidermal tissue, and may help the development of antiviral measures. Human papillomaviruses (HPVs) comprise a large family of small, nonenveloped DNA viruses with transforming potential. HPVs selectively infect basal keratinocytes of stratified skin and mucosal epithelia and persist, mostly without clinical symptoms, in virtually every part of the human skin. The biological costs of HPV persistence range from benign papilloma and genital warts over preneoplastic lesions to anogenital or oropharyngeal cancers (1). In fact, infection by the so-called "high-risk" HPV causes about 5% of all human cancers (2). Of these, cervical cancers are the most prevalent. However, HPV-associated oropharyngea...

show abstract

“…After endocytosis and vesicle acidification, capsid disassembly occurs within the endosome, thereby releasing a complex of viral genome and the L2 protein. The minor capsid protein mediates all further steps essential for establishing infection (20,(28)(29)(30)(31)(32)(33).…”

mentioning

confidence: 99%

Tetraspanin CD151 Mediates Papillomavirus Type 16 Endocytosis

Scheffer

Gawlitza

Spoden

et al. 2013

J Virol

118

171

View full text Add to dashboard Cite

cHuman papillomavirus type 16 (HPV16) is the primary etiologic agent for cervical cancer. The infectious entry of HPV16 into cells occurs via a so-far poorly characterized clathrin-and caveolin-independent endocytic pathway, which involves tetraspanin proteins and actin. In this study, we investigated the specific role of the tetraspanin CD151 in the early steps of HPV16 infection. We show that surface-bound HPV16 moves together with CD151 within the plane of the membrane before they cointernalize into endosomes. Depletion of endogenous CD151 did not affect binding of viral particles to cells but resulted in reduction of HPV16 endocytosis. HPV16 uptake is dependent on the C-terminal cytoplasmic region of CD151 but does not require its tyrosine-based sorting motif. Reexpression of the wild-type CD151 but not mutants affecting integrin functions restored virus internalization in CD151-depleted cells. Accordingly, short interfering RNA (siRNA) gene knockdown experiments confirmed that CD151-associated integrins (i.e., ␣3␤1 and ␣6␤1/4) are involved in HPV16 infection. Furthermore, palmitoylation-deficient CD151 did not support HPV16 cell entry. These data show that complex formation of CD151 with laminin-binding integrins and integration of the complex into tetraspanin-enriched microdomains are critical for HPV16 endocytosis.

show abstract

Principles of polyoma- and papillomavirus uncoating

Cited by 17 publications

References 131 publications

Translocation of the papillomavirus L2/vDNA complex across the limiting membrane requires the onset of mitosis

Translocation of the papillomavirus L2/vDNA complex across the limiting membrane requires the onset of mitosis

Kallikrein-8 Proteolytically Processes Human Papillomaviruses in the Extracellular Space To Facilitate Entry into Host Cells

Tetraspanin CD151 Mediates Papillomavirus Type 16 Endocytosis

Contact Info

Product

Resources

About