The Lassa virus glycoprotein precursor GP-C is proteolytically processed by subtilase SKI-1/S1P

Lenz, Oliver; Meulen, Jan ter; Klenk, Hans‐Dieter; Seidah, Nabil G.; Garten, Wolfgang

doi:10.1073/pnas.221447598

Cited by 331 publications

(383 citation statements)

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“…2-4). It may well be that the RRLL sequence that is best recognized by SKI-1 in a number of substrates (7,44,48) may actually be cleaved when present in the RSL of ␣ 1 -AT. Thus, whereas only an RSL with a P1 Leu in combination with either P2 Val, Ile, and Tyr resulted in an inhibitory serpin, we cannot eliminate the possibility that a P1 Val or Ile with a P2 Val, Ile, or Tyr may not also be inhibitory.…”

Section: Discussionmentioning

confidence: 99%

“…In the ␣ 1 -AT variants, we mutated the reactive site loop sequence AIPM 358 (P1-P4 positions) to mimic the reported SKI-1 specificity for the motif (R/K)X(hydrophobic)Z2 (5-7), with a preference for the RRLL2 cleavage site reported for the SKI-1 prosegment site C (5), and the glycoproteins of Lassa virus (44,45) and CCHV virus (48). Co-expression of pro-PDGF-A or pro-PDGF-A* with either ␣ 1 -AT, ␣ 1 -PDX, or various serpin mutants are shown in Fig.…”

Section: Inhibition Of Pro-pdgf-a* (Rrll 86 ) Processing By Ski-1 In mentioning

confidence: 99%

“…Mutation of proplatelet-derived growth factor A (pro-PDGF-A) at its furin-cleavage site (RRKR 86 ) into RRLL 86 (pro-PDGF-A*) resulted in a SKI-1 artificial substrate (43). Finally, SKI-1 was shown to play a major role in the processing of surface glycoproteins of infectious viruses such as Lassa (44,45), lymphocytic choriomeningitis (LCMV) (46,47) and Crimean Congo hemorrhagic fever (CCHF) (48) viruses. CCHF is a tick-borne member of the genus Nairovirus, family Bunyaviridae.…”

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Development of Protein-based Inhibitors of the Proprotein of Convertase SKI-1/S1P

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Vincent

Nichol

et al. 2004

Journal of Biological Chemistry

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Processing of membrane-bound transcription factors such as sterol regulatory element-binding proteins (SREBPs) and the ER-stress response factor ATF6, and glycoproteins of some hemorrhagic fever viruses are initiated by the proprotein convertase SKI-1/S1P. So far, no cellular protein-based inhibitor of the hydrophobicamino acid specific SKI-1 is known. The prosegment of the basic-amino acid specific convertases (e.g. furin and PC5) or ␣ 1 -PDX, a variant of ␣ 1 -antitrypsin (␣ 1 -AT) exhibiting an RIPR 358 sequence at the reactive site loop, were shown to potently inhibit these secretory proteinases. Accordingly, we tested the SKI-1-inhibitory potential of various point mutants of either the 198 amino acid preprosegment of SKI-1-(1-198) or ␣ 1 -AT. Transient transfections data showed that, out of numerous mutants studied, the R134E prosegment mutant or the ␣ 1 -AT reactive site loop variants RRVL 358 , RRYL 358 and RRIL 358 are the best specific cellular inhibitors of SKI-1. The observed inhibition of the processing of endogenous SREBP-2, exogenous ATF6 and a PDGF-A (RRLL 86 ) variant were >55% and reach ϳ80% in stable transfectants. We also show that SKI-1 forms SDS-stable complexes with these ␣ 1 -AT variants, but not with wild-type ␣ 1 -AT or ␣ 1 -PDX. Finally, these inhibitors were also shown to affect the processing and stability of the Crimean-Congo hemorrhagic fever virus glycoprotein.Proteins and peptides that are biologically active are often generated by intracellular limited proteolysis of inactive precursors. The mammalian proprotein convertases (PCs) 1 of the secretory pathway are calcium-dependent serine proteinases related to bacterial subtilisin that cleave various precursors at the general consensus motif (K/R)(X) n (K/R)2, where n ϭ 0, 2, 4, or 6 and X is any amino acid (1-3). The PC family counts seven basic amino acid-specific kexin-like convertases: furin, PC1/3, PC2, PC4, PACE4, PC5/6, and PC7/LPC (4). The eighth member is the recently discovered pyrolysin-like SKI-1/S1P that cleaves at the consensus motif (R/K)X(hydrophobic)Z2, where Z is variable (5), while the last member (6, 7) NARC-1 cleaves the sequence VFAQ 153 2 in its prosegment (8, 9). 2 More PCs contain an N-terminal signal sequence, followed by a prosegment, a catalytic domain and a P-domain. In addition, PCs possess a C-terminal segment that varies between the different members. The critical role of PCs in the proteolytic maturation of multiple proproteins, their implication in various pathologies (1, 10, 11), and their unidentified specific and/or redundant functions, make them attractive targets for the development of potent and selective inhibitors. The various successful approaches include: active site-directed chloromethyl ketone inhibitors (12, 13), reversible peptide-based inhibitors (14 -17), plant derivatives (18), and several engineered variants of protein-based inhibitors that possess a furin-like motif. These include ␣2-macroglobulin (␣2-MF) (19), ␣ 1 -antitrypsin (␣ 1 -AT) Portland (␣ 1 -PDX) (20 -22), proteinase i...

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Section: Discussionmentioning

confidence: 99%

Section: Inhibition Of Pro-pdgf-a* (Rrll 86 ) Processing By Ski-1 In mentioning

confidence: 99%

mentioning

confidence: 99%

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Development of Protein-based Inhibitors of the Proprotein of Convertase SKI-1/S1P

Pullikotil

Vincent

Nichol

et al. 2004

Journal of Biological Chemistry

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“…The WE strain of LCMV was kindly provided by D. von Laer, Innsbruck, Austria. [5]. Cells were transfected with plasmids containing cDNA of the viral glycoprotein constructs using Lipofectamine 2000 (Invitrogen) according to manufacturer's instruction.…”

Section: Molecular Cloning and Protein Expressionmentioning

confidence: 99%

“…The glycoprotein of LASV is synthesized as the precursor molecule preGP-C which is co-translationally processed by the cellular signal peptidase resulting in the 6 kDa stable signal peptide (SSP) and the immature 76 kDa GP-C. Uncommonly, after cleavage the SSP stays connected to the glycoprotein complex [2][3][4]. During the transport along the secretory pathway to the plasma membrane, GP-C is then further proteolytically activated by maturation cleavage into the distal glycoprotein subunit (GP-1) (44 kDa) and the transmembrane-spanning subunit (GP-2) (36 kDa) by the cellular endoprotease subtilase subtilisin kexin isozyme-1/site 1 protease (SKI-1/S1P) [5,6]. During this cleavage step, the SSP acts as an essential trans-acting maturation factor [3,7].…”

Section: Introductionmentioning

confidence: 99%

Maturation cleavage within the ectodomain of Lassa virus glycoprotein relies on stabilization by the cytoplasmic tail

2010

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a b s t r a c tThe Lassa virus glycoprotein consists of an ectodomain, a transmembrane anchor, and a cytoplasmic domain. It is synthesized as an inactive precursor and cleaved within the ectodomain to yield the mature form. Here, we show that this maturation cleavage can be abolished by mutation of single conserved amino acids within the cytoplasmic domain at the carboxy-terminus of the glycoprotein. Moreover, substitutions and deletions of multiple amino acids result in destabilization of the glycoprotein oligomers. These results indicate that conformation changes in the cytoplasmic domain travel across the membrane and subsequently abolish the maturation cleavage. Therefore, we postulate that the cytoplasmic domain is an important maturation factor stabilizing the overall conformation of the glycoprotein. Structured summary:MINT-7997004: LASV GP (uniprotkb:P08669) and LASV GP (uniprotkb:P08669) physically interact (MI:0915) by cosedimentation through density gradient (MI:0029)

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Arenaviruses

Kunz¹

2009

Encyclopedia of Life Sciences

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The arenaviruses are a large and diverse group of enveloped ribonucleic acid (RNA) viruses that merit significant attention as powerful experimental models and important human pathogens. The prototypic arenavirus lymphocytic choriomeningitis virus (LCMV) has been instrumental in many landmark studies on virus–host interaction and antiviral immunity. Several arenaviruses, including Lassa virus in Africa and the South American arenaviruses such as Junin virus , Guanarito virus , Machupo virus and Sabia virus cause severe viral haemorrhagic fevers in humans and represent serious public health problems. Arenavirus haemorrhagic fevers are among the most devastating emerging human diseases with high fatality rates in hospitalized patients and a limited therapeutic repertoire available. Moreover, compelling evidence indicates that the worldwide‐distributed LCMV is a neglected human pathogen of clinical significance, especially in cases of congenital infection and immunocompromised individuals, as tragically illustrated by recent fatal cases of transplant‐associated infections. Key concepts Arenaviruses are zoonotic pathogens that are carried in nature by persistent infection of rodent hosts. The interaction of prototypic arenavirus LCMV with its host species, the mouse, provided fundamental concepts in virology and immunology. The current diversity of arenaviruses is the result of a remarkable long‐term co‐evolution between the viruses and their rodent hosts. When infecting humans as accidental hosts, some arenaviruses cause severe haemorrhagic fevers with high mortality. Arenavirus haemorrhagic fevers belong to the worst human diseases and represent important public health problems in many regions of the developing world. The epidemiology, clinical disease, diagnostics and current treatment for these diseases will be discussed. During the past decade much has been learned about the molecular and cell biology of the arenavirus life cycle. Cellular receptors for all major human pathogenic arenaviruses have been identified and first light has been shed on their mechanisms of cell entry. The advent of a powerful reverse genetic system for arenaviruses has allowed to elucidate the mechanisms of arenavirus replication, gene expression and assembly and release of progeny virions from infected cells. The investigation of the fundamental virology of arenaviruses will likely provide novel avenues for the development of efficacious therapeutic strategies against these important human pathogens.

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The Lassa virus glycoprotein precursor GP-C is proteolytically processed by subtilase SKI-1/S1P

Cited by 331 publications

References 28 publications

Development of Protein-based Inhibitors of the Proprotein of Convertase SKI-1/S1P

Development of Protein-based Inhibitors of the Proprotein of Convertase SKI-1/S1P

Maturation cleavage within the ectodomain of Lassa virus glycoprotein relies on stabilization by the cytoplasmic tail

Arenaviruses

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