Unexpected Tolerance of α-Cleavage of the Prion Protein to Sequence Variations

Oliveira-Martins, José B.; Yusa, Sei-ichi; Calella, Anna Maria; Bridel, Claire; Baumann, Frank; Dametto, Paolo; Aguzzi, Adriano

doi:10.1371/journal.pone.0009107

Cited by 44 publications

(60 citation statements)

References 53 publications

(107 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…However, it is also to be considered whether the original formulation of the π hypothesis is compatible with our current knowledge of PrP C chemistry. For example, the π hypothesis envisages two discrete, covalently linked binding sites on PrP C (one N terminal, one C terminal) simultaneously docking onto the L PrP protein, yet we now know that much of PrP C at steady-state is endoproteolysed to separate the N-and C-terminal domains (31,(41)(42)(43). Furthermore, other hypotheses that also seek to explain the properties of internal PrP deletions neither invoke a PrP paralog nor consider action in embryonic development (44).…”

Section: Activities Needed To Maintain Cell Viability In the Adult Cnmentioning

confidence: 99%

Knockout of the prion protein (PrP)-like Sprn gene does not produce embryonic lethality in combination with PrP ^C -deficiency

Daude¹,

Wohlgemuth²,

Brown

et al. 2012

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

View full text Add to dashboard Cite

The Sprn gene encodes Shadoo (Sho), a glycoprotein with biochemical properties similar to the unstructured region of cellular prion protein (PrP C ). Sho has been considered a candidate for the hypothetical π protein that supplies a PrP C -like function to maintain the viability of Prnp 0/0 mice lacking the PrP C protein. To understand these relationships more clearly we probed the cell biology of Sho and created knockout mice. Besides full-length and a “C1” C-terminal fragment, we describe a 6-kDa N-terminal Sho neuropeptide, “N1,” which is present in membrane-enriched subcellular fractions of wild-type mice. Sprn null alleles were produced that delete all protein coding sequences yet spare the Mtg 1 gene transcription unit that overlaps the Sprn 3′ UTR; the resulting mice bred to homozygosity were viable and fertile, although Sprn 0/0 mice maintained in two genetic backgrounds weighed less than wild-type mice. Lack of Sho protein did not affect prion incubation time. Contrasting with lethality reported for knockdown of expression in Prnp 0/0 embryos using lentiviruses targeted against the Sprn 3′ UTR, we established that double-knockout mice deficient in both Sho and PrP C are fertile and viable up to 690 d of age. Our data reduce the impetus for equating Sho with the notional π protein and are not readily reconciled with hypotheses wherein expression of PrP C and Sho are both required for completion of embryogenesis. Alternatively, and in accord with some reports for PrP C , we infer that Sho’s activity will prove germane to the maintenance of neuronal viability in postnatal life.

show abstract

Section: Activities Needed To Maintain Cell Viability In the Adult Cnmentioning

confidence: 99%

Knockout of the prion protein (PrP)-like Sprn gene does not produce embryonic lethality in combination with PrP ^C -deficiency

Daude¹,

Wohlgemuth²,

Brown

et al. 2012

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

View full text Add to dashboard Cite

show abstract

“…The HD is essential for PrP C homodimerization and ␣-cleavage (Rambold et al, 2008;Bremer et al, 2010;Oliveira-Martins et al, 2010). We sought to directly test whether PrP C homodimerization could modulate the ␣-cleavage.…”

Section: Prp C Homodimerization Increases Prpc1 and Prpn1 Levels In Cmentioning

confidence: 99%

“…Dimerization-mediated ␣-cleavage of PrP C is independent of ADAM8, 10, and 17 The two metalloproteases ADAM10 and ADAM17 were initially proposed to be the proteases that are responsible for constitutive and PKC-regulated ␣-cleavage of PrP C , respectively, yet their involvement is currently a matter of debate (Vincent et al, 2001;Taylor et al, 2009;Oliveira-Martins et al, 2010;Altmeppen et al, 2011). Recent results suggest that ADAM8 regulates PrP C ␣-cleavage in skeletal muscle (Liang et al, 2012).…”

Section: Homodimerization Increases Prpmentioning

confidence: 99%

“…Overall, these studies point to a possible neuroprotective activity associated with ␣-cleavage of PrP C . In favor of this hypothesis, PrP Sc and toxic PrP mutants are impaired in their ␣-cleavage (Oliveira- Martins et al, 2010).…”

Section: Introductionmentioning

confidence: 99%

“…However, ␣-cleavage occurs normally in neuron-specific ADAM10 knock-out mice and the search for other proteases that have the ability to perform the ␣-cleavage continues (Altmeppen et al, 2011). The hydrophobic domain (HD) (amino acids 111-129) of PrP C is essential for the ␣-cleavage both in cellulo and in vivo (Bremer et al, 2010;Oliveira-Martins et al, 2010). Interestingly, the HD is also essential for PrP C homodimerization and the stress-protective activity of PrP C (Rambold et al, 2008).…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

PrP^CHomodimerization Stimulates the Production of PrP^CCleaved Fragments PrPN1 and PrPC1

et al. 2012

View full text Add to dashboard Cite

An endoproteolytic cleavage termed ␣-cleavage between residues 111/112 is a characteristic feature of the cellular prion protein (PrP C ). This cleavage generates a soluble N-terminal fragment (PrPN1) and a glycosylphosphatidylinositol-anchored C-terminal fragment (PrPC1). Independent studies demonstrate that modulating PrP C ␣-cleavage represents a potential therapeutic strategy in prion diseases. The regulation of PrP C ␣-cleavage is unclear. The only known domain that is essential for the ␣-cleavage to occur is a hydrophobic domain (HD). Importantly, the HD is also essential for the formation of PrP C homodimers. To explore the role of PrP C homodimerization on the ␣-cleavage, we used a well described inducible dimerization strategy whereby a chimeric PrP C composed of a modified FK506-binding protein (Fv) fused with PrP C and termed Fv-PrP is incubated in the presence of a dimerizer AP20187 ligand. We show that homodimerization leads to a considerable increase of PrP C ␣-cleavage in cultured cells and release of PrPN1 and PrPC1. Interestingly, enforced homodimerization increased PrP C levels at the plasma membrane, and preventing PrP C trafficking to the cell surface inhibited dimerization-induced ␣-cleavage. These observations were confirmed in primary hippocampal neurons from transgenic mice expressing Fv-PrP. The proteases responsible for the ␣-cleavage are still elusive, and in contrast to initial studies we confirm more recent investigations that neither ADAM10 nor ADAM17 are involved. Importantly, PrPN1 produced after PrP C homodimerization protects against toxic amyloid-␤ (A␤) oligomers. Thus, our results show that PrP C homodimerization is an important regulator of PrP C ␣-cleavage and may represent a potential therapeutic avenue against A␤ toxicity in Alzheimer's disease.

show abstract

Effects of FlAsH/Tetracysteine (TC) Tag on PrP Proteolysis and PrPres Formation by TC‐Scanning

Taguchi¹,

Hohsfield²,

Hollister³

et al. 2013

ChemBioChem

View full text Add to dashboard Cite

The FlAsH/tetracysteine (FlAsH/TC) tag is a powerful tool for fluorescent labeling of proteins. However, even small tags such as FlAsH/TC could alter the behavior of the tagged proteins, especially if the insertion occurs at internal sites. Defining the influence of FlAsH/TC on nearby protein-protein interactions might aid in selecting appropriate positions for internal TC insertions and allow the exploitation of serial FlAsH/TC insertions (TC-scanning) as a probe to characterize sites of protein-protein interaction. To explore this application in the context of substrate-protease interactions, we analyzed the effect of FlAsH/TC insertions on proteolysis of cellular prion protein (PrPsen) in in vitro reactions and generation of the C1 metabolic fragment of PrPsen in live neuroblastoma cells. The influence of FlAsH/TC insertion was evaluated by TC-scanning across the cleavage sites of each protease. The results showed that FlAsH/TC inhibited protease cleavage only within limited ranges of the cleavage sites that varied from about 1 to 6 residues-wide depending on the protease, providing an estimate of the PrP residues interacting with each protease. TC-scanning was also used to probe a different type of protein-protein interaction, the conformational conversion of FlAsH-PrPsen to the prion disease-associated isoform, PrPres. PrP constructs with FlAsH/TC insertions at residues 90–96 but not 97–101 were converted to FlAsH-PrPres, identifying a boundary separating loosely versus compactly folded regions of PrPres. Our observations demonstrate that TC-scanning with the FlAsH/TC tag can be a versatile method for probing protein-protein interactions and folding processes.

show abstract

Unexpected Tolerance of α-Cleavage of the Prion Protein to Sequence Variations

Cited by 44 publications

References 53 publications

Knockout of the prion protein (PrP)-like Sprn gene does not produce embryonic lethality in combination with PrP ^C -deficiency

Knockout of the prion protein (PrP)-like Sprn gene does not produce embryonic lethality in combination with PrP ^C -deficiency

PrP^CHomodimerization Stimulates the Production of PrP^CCleaved Fragments PrPN1 and PrPC1

Effects of FlAsH/Tetracysteine (TC) Tag on PrP Proteolysis and PrPres Formation by TC‐Scanning

Contact Info

Product

Resources

About

Unexpected Tolerance of α-Cleavage of the Prion Protein to Sequence Variations

Cited by 44 publications

References 53 publications

Knockout of the prion protein (PrP)-like Sprn gene does not produce embryonic lethality in combination with PrP C -deficiency

Knockout of the prion protein (PrP)-like Sprn gene does not produce embryonic lethality in combination with PrP C -deficiency

PrPCHomodimerization Stimulates the Production of PrPCCleaved Fragments PrPN1 and PrPC1

Effects of FlAsH/Tetracysteine (TC) Tag on PrP Proteolysis and PrPres Formation by TC‐Scanning

Contact Info

Product

Resources

About

Knockout of the prion protein (PrP)-like Sprn gene does not produce embryonic lethality in combination with PrP ^C -deficiency

Knockout of the prion protein (PrP)-like Sprn gene does not produce embryonic lethality in combination with PrP ^C -deficiency

PrP^CHomodimerization Stimulates the Production of PrP^CCleaved Fragments PrPN1 and PrPC1