Ectodomain shedding of furin: kinetics and role of the cysteine‐rich region

Denault, Jean‐Bernard; Bissonnette, Lyne; Longpré, Jean‐Michel; Charest, Gabriel; Lavigne, Pierre; Leduc, Richard

doi:10.1016/s0014-5793(02)03249-0

Cited by 28 publications

(33 citation statements)

References 43 publications

(49 reference statements)

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“…S1 C and D). Thus, PC5/6 was preferentially associated with the BM, while furin was highly expressed on basal cells, from which it might be shed or secreted to act on virions bound to the BM (12,13).…”

Section: Furin Inhibition Decreases In Vivo Infection Through Loss Ofmentioning

confidence: 99%

The initial steps leading to papillomavirus infection occur on the basement membrane prior to cell surface binding

Kines

Thompson

Lowy

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

236

230

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Using a murine challenge model, we previously determined that human papillomavirus (HPV) pseudovirions initially bind preferentially to the cervicovaginal basement membrane (BM) at sites of trauma. We now report that the capsids undergo a conformational change while bound to the BM that results in L2 cleavage by a proprotein convertase (PC), furin, and/or PC5/6, followed by the exposure of an N-terminal cross-neutralization L2 epitope and transfer of the capsids to the epithelial cell surface. Prevention of this exposure by PC inhibition results in detachment of the pseudovirions from the BM and their eventual loss from the tissue, thereby preventing infection. Pseudovirions whose L2 had been precleaved by furin can bypass the PC inhibition of binding and infectivity. Cleavage of heparan sulfate proteoglycans (HSPG) with heparinase III prevented infection and BM binding by the precleaved pseudovirions, but did not prevent them from binding robustly to cell surfaces. These results indicate that the infectious process has evolved so that the initial steps take place on the BM, in contrast to the typical viral infection that is initiated by binding to the cell surface. The data are consistent with a dynamic model of in vivo HPV infection in which a conformational change and PC cleavage on the BM allows transfer of virions from HSPG attachment factors to an L1-specific receptor on basal keratinocytes migrating into the site of trauma.furin ͉ HSPG ͉ mouse model ͉ proprotein convertase P apillomaviruses (PVs) infect mucosal and cutaneous stratified squamous epithelia, resulting in benign lesions, some of which can progress to invasive cancer. Human papillomavirus (HPV) 16 and several other PVs that preferentially infect the anogenital mucosa are responsible for virtually all cases of cervical cancer, as well as a number of other mucosal epithelial malignancies. It has been recognized for decades that efficient induction of experimental PV infection of animals requires local wounding in conjunction with virus exposure (1-4). However, there has been little molecular insight into the in vivo steps by which PVs initiate the infectious process.We recently described a murine cervicovaginal challenge model of PV transmission that is amenable to examination of the early in vivo events of PV infection (5, 6). It employs high titer PV pseudoviruses, which are authentic PV capsids composed of the L1 major and L2 minor structural proteins that have encapsidated a non-PV plasmid encoding a quantifiable reporter gene to monitor successful infection (7). PV pseudoviruses abrogate the exquisite species specificity of the viral gene expression program of authentic PV virions, while retaining the early events of PV infection, including the preferential infection of keratinocytes in vivo. In the initial characterization of the mouse genital tract model, we confirmed that wounding by the transient disruption of cervicovaginal epithelial integrity, by physical or chemical means, is essential for efficient infection (5). It was shown that ...

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Section: Furin Inhibition Decreases In Vivo Infection Through Loss Ofmentioning

confidence: 99%

The initial steps leading to papillomavirus infection occur on the basement membrane prior to cell surface binding

Kines

Thompson

Lowy

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

236

230

View full text Add to dashboard Cite

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“…In contrast, Furin has a transmembrane domain and binds cytosolic adapters that mediate cycling between the trans-Golgi network, the cell surface, and endosomes (Thomas 2002). The acidic pH of the trans-Golgi network/endosomal system favors a second autocatalytic cleavage displacing an inhibitory propeptide (Anderson et al 2002), and cleavage of the membrane anchor by an unknown ''sheddase'' in acidic compartments can give rise to a soluble form of activated Furin (Wise et al 1990;Vidricaire et al 1993;Denault et al 2002). Shed Furin has been observed in the milk of transgenic mice (Paleyanda et al 1997) and in epididymal fluid (Thimon et al 2006).…”

mentioning

confidence: 99%

The microenvironment patterns the pluripotent mouse epiblast through paracrine Furin and Pace4 proteolytic activities

Mesnard¹,

Donnison²,

Fuerer³

et al. 2011

Genes Dev.

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The fate of pluripotent cells in early mouse embryos is controlled by graded Nodal signals that are activated by the endoproteases Furin and Pace4. Soluble forms of Furin and Pace4 cleave proNodal in vitro and after secretion in transfected cells, but direct evidence for paracrine activity in vivo is elusive. Here, we show that Furin and Pace4 are released by the extraembryonic microenvironment, and that they cleave a membrane-bound reporter substrate in adjacent epiblast cells and activate Nodal to maintain pluripotency. Secreted Pace4 and Furin also stimulated mesoderm formation, whereas endoderm was only induced by Pace4, correlating with a difference in the spatiotemporal distribution of these proteolytic activities. Our analysis of paracrine Furin and Pace4 activities and their in vivo functions significantly advances our understanding of how the epiblast is patterned by its microenvironment. Adding cell-cell communication to the pleiotropic portfolio of these proteases provides a new framework to study proprotein processing also in other relevant contexts.

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“…It has not yet been clearly established whether the C-terminal cleavage of furin occurs intracellularly, leading to the secretion of the truncated form, or whether it occurs once the mature protein is on the cell surface. In the former case, it is suggested that furin itself or a related PC is involved in this process (Denault et al 2002), although the cleavage site is not a canonical sequence for these enzymes. Moreover, the fact that mutation or deletion of the cleavage site severely reduces ectodomain release without suppressing it suggests that there is no definite cleavage site (Plaimauer et al 2001).…”

Section: Discussionmentioning

confidence: 99%

Analysis of furin ectodomain shedding in epididymal fluid of mammals: demonstration that shedding of furin occurs in vivo

Thimon¹,

Belghazi²,

Dacheux³

et al. 2006

Reproduction

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Sperm cell surface proteins and proteins of their surrounding fluids are reported to be proteolytically processed in relation to acquisition of sperm fertility during epididymal transit. Several of these proteins might be potential targets for subtilisin-like proprotein convertase. Using immunochemistry and mass spectrometry analysis, we found that an 80 kDa form of furin (EC 3.4.21.75) is present in the fluid from the mid-caput to the distal corpus regions of the epididymis of various domestic mammals. This protein is absent from the fluid of the caudal region, suggesting that it is reabsorbed or degraded. The cDNA sequence of ovine furin was obtained and the mRNA was found throughout this organ, although in greater amounts in the mid and distal caput regions. Metabolic labeling with 35 S-amino acids indicated that the protein was synthesized and released from the epithelium only in a restricted area of the mid-caput, suggesting a specific regionalized mechanism of secretion. The fluid protein is not pelleted at 100 000 g and did not react with a C-terminal antibody indicating that it is not bound to membranous materials. These findings demonstrate that a furin ectodomain shedding occurs naturally in vivo in the epididymis where this enzyme could be involved in fluid and/or sperm membrane protein processing.Reproduction (2006) 132 899-908

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Ectodomain shedding of furin: kinetics and role of the cysteine‐rich region

Cited by 28 publications

References 43 publications

The initial steps leading to papillomavirus infection occur on the basement membrane prior to cell surface binding

The initial steps leading to papillomavirus infection occur on the basement membrane prior to cell surface binding

The microenvironment patterns the pluripotent mouse epiblast through paracrine Furin and Pace4 proteolytic activities

Analysis of furin ectodomain shedding in epididymal fluid of mammals: demonstration that shedding of furin occurs in vivo

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