Proteolytic Cleavage of Bovine Adenovirus 3-Encoded pVIII

ABSTRACT Proteolytic maturation involving cleavage of one nonstructural and six structural precursor proteins including pVIII by adenovirus protease is an important aspect of the adenovirus life cycle. The pVIII encoded by bovine adenovirus 3 (BAdV-3) is a protein of 216 amino acids and contains two potential protease cleavage sites. Here, we report that BAdV-3 pVIII is cleaved by adenovirus protease at both potential consensus protease cleavage sites. Usage of at least one cleavage site appears essential for the production of progeny BAdV-3 virions as glycine-to-alanine mutation of both protease cleavage sites appears lethal for the production of progeny virions. However, mutation of a single protease cleavage site of BAdV-3 pVIII significantly affects the efficient production of infectious progeny virions. Further analysis revealed no significant defect in endosome escape, genome replication, capsid formation, and virus assembly. Interestingly, cleavage of pVIII at both potential cleavage sites appears essential for the production of stable BAdV-3 virions as BAdV-3 expressing pVIII containing a glycine-to-alanine mutation of either of the potential cleavage sites is thermolabile, and this mutation leads to the production of noninfectious virions. IMPORTANCE Here, we demonstrated that the BAdV-3 adenovirus protease cleaves BAdV-3 pVIII at both potential protease cleavage sites. Although cleavage of pVIII at one of the two adenoviral protease cleavage sites is required for the production of progeny virions, the mutation of a single cleavage site of pVIII affects the efficient production of infectious progeny virions. Further analysis indicated that the mutation of a single protease cleavage site (glycine to alanine) of pVIII produces thermolabile virions, which leads to the production of noninfectious virions with disrupted capsids. We thus provide evidence about the requirement of proteolytic cleavage of pVIII for production of infectious progeny virions. We feel that our study has significantly advanced the understanding of the requirement of adenovirus protease cleavage of pVIII.

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Human Immunodeficiency Virus Type 1 Protease Cleavage Site Mutations Associated with Protease Inhibitor Cross-Resistance Selected by Indinavir, Ritonavir, and/or Saquinavir

Journal of Virology ◽

10.1128/jvi.75.2.589-594.2001 ◽

2001 ◽

Vol 75 (2) ◽

pp. 589-594 ◽

Cited By ~ 92

Author(s):

Hélène C. F. Côté ◽

Zabrina L. Brumme ◽

P. Richard Harrigan

Keyword(s):

Human Immunodeficiency Virus ◽

Protease Inhibitor ◽

Cleavage Site ◽

Cross Resistance ◽

Cleavage Sites ◽

Protease Cleavage ◽

Immunodeficiency Virus ◽

Before And After ◽

Protease Cleavage Sites

ABSTRACT We examined the prevalence of cleavage site mutations, both within and outside the gag region, in 28 protease inhibitor (PI) cross-resistant patients treated with indinavir, ritonavir, and/or saquinavir compared to control patients treated with reverse transcriptase inhibitors. Three human immunodeficiency virus protease cleavage sites within gag (p2/NC, NC/p1, and NC/TFP) showed considerable in vivo evolution before and after therapy with indinavir, ritonavir, and/or saquinavir. Another gag cleavage site (p1/p6 gag ) showed a trend compared to matched controls. The other eight recognized cleavage sites showed relatively little difference between PI-resistant cases and controls. An A→V substitution at the P2 position of the NC/p1 and NC/TFP cleavage sites was the most common (29%) change selected by the PIs used in this study.

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Ty1 Proteolytic Cleavage Sites Are Required for Transposition: All Sites Are Not Created Equal

Journal of Virology ◽

10.1128/jvi.75.2.638-644.2001 ◽

2001 ◽

Vol 75 (2) ◽

pp. 638-644 ◽

Cited By ~ 22

Author(s):

Gennady V. Merkulov ◽

Joseph F. Lawler ◽

Yolanda Eby ◽

Jef D. Boeke

Keyword(s):

Virus Assembly ◽

The Other ◽

Cleavage Sites ◽

Protease Cleavage ◽

Sequence Of Events ◽

Key Enzyme ◽

Protease Cleavage Sites ◽

Proteolytic Cleavages ◽

Retroviral Protease

ABSTRACT The retroviral protease is a key enzyme in a viral multienzyme complex that initiates an ordered sequence of events leading to virus assembly and propagation. Viral peptides are initially synthesized as polyprotein precursors; these precursors undergo a number of proteolytic cleavages executed by the protease in a specific and presumably ordered manner. To determine the role of individual protease cleavage sites in Ty1, a retrotransposon from Saccharomyces cerevisiae, the cleavage sites were systematically mutagenized. Altering the cleavage sites of the yeast Ty1 retrotransposon produces mutants with distinct retrotransposition phenotypes. Blocking the Gag/PR site also blocks cleavage at the other two cleavage sites, PR/IN and IN/RT. In contrast, mutational block of the PR/IN or IN/RT sites does not prevent cleavage at the other two sites. Retrotransposons with mutations in each of these sites have transposition defects. Mutations in the PR/IN and IN/RT sites, but not in the Gag/PR site, can be complemented in trans by endogenous Ty1 copies. Hence, the digestion of the Gag/PR site and release of the protease N terminus is a prerequisite for processing at the remaining sites; cleavage of PR/IN is not required for the cleavage of IN/RT, and vice versa. Of the three cleavage sites in the Gag-Pol precursor, the Gag/PR site is processed first. Thus, Ty1 Gag-Pol processing proceeds by an ordered pathway.

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Family-wide characterization of matrix metalloproteinases from Arabidopsis thaliana reveals their distinct proteolytic activity and cleavage site specificity

Biochemical Journal ◽

10.1042/bj20130196 ◽

2013 ◽

Vol 457 (2) ◽

pp. 335-346 ◽

Cited By ~ 23

Author(s):

Giada Marino ◽

Pitter F. Huesgen ◽

Ulrich Eckhard ◽

Christopher M. Overall ◽

Wolfgang P. Schröder ◽

...

Keyword(s):

Arabidopsis Thaliana ◽

Cleavage Site ◽

Biochemical Properties ◽

Site Specificity ◽

Sequence Specificity ◽

Cleavage Sites ◽

Protease Cleavage ◽

Protease Cleavage Sites ◽

Proteomic Identification

The five recombinant MMP-like proteins of Arabidopsis thaliana have specific biochemical properties. Detailed analysis of their sequence specificity using proteomic identification of protease cleavage sites revealed cleavage profiles similar to human MMPs.

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A “molecular guillotine” reveals an interphase function of Kinesin-5

10.1101/184416 ◽

2017 ◽

Author(s):

Zhiyi Lv ◽

Jan Rosenbaum ◽

Timo Aspelmeier ◽

Jörg Großhans

Keyword(s):

Cleavage Site ◽

Motor Proteins ◽

Motor Protein ◽

Proteolytic Cleavage ◽

General Strategy ◽

Microtubule Network ◽

Tev Protease ◽

Protease Cleavage Site ◽

Protease Cleavage ◽

Drosophila Embryos

AbstractMotor proteins are important for transport and force generation in a variety of cellular processes and morphogenesis. Here we design a general strategy for conditional motor mutants by inserting a protease cleavage site at the “neck” between the head domain and the stalk of the motor protein, making the protein susceptible to proteolytic cleavage at the neck by the corresponding protease. To demonstrate the feasibility of this approach, we inserted the cleavage site of TEV protease into the neck of the tetrameric motor Kinesin-5. Application of TEV protease led to a specific depletion and functional loss of Kinesin-5 in Drosophila embryos. By this, we revealed that Kinesin-5 stabilized the microtubule network during interphase in syncytial embryos. The “molecular guillotine” can potentially be applied to many motor proteins due to the conserved structures of kinesin, dynein and myosin with accessible necks.Author summaryWe design a general strategy for conditional motor mutants by inserting a protease cleavage site between head and stalk domain of the motor protein, making it susceptible to specific proteolytic cleavage. We demonstrate the feasibility of the approach with the motor Kinesin-5 and the protease TEV in Drosophila embryos. This approach can potentially be applied to motor proteins kinesin, dynein and myosin due to the conserved structures.

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Variability at Human Immunodeficiency Virus Type 1 Subtype C Protease Cleavage Sites: an Indication of Viral Fitness?

Journal of Virology ◽

10.1128/jvi.77.17.9422-9430.2003 ◽

2003 ◽

Vol 77 (17) ◽

pp. 9422-9430 ◽

Cited By ~ 45

Author(s):

Tulio de Oliveira ◽

Susan Engelbrecht ◽

Estrelita Janse van Rensburg ◽

Michelle Gordon ◽

Karen Bishop ◽

...

Keyword(s):

Human Immunodeficiency Virus ◽

Human Immunodeficiency Virus Type ◽

Cleavage Site ◽

Cleavage Sites ◽

Subtype C ◽

Protease Cleavage ◽

Immunodeficiency Virus ◽

Protease Cleavage Sites ◽

Hiv 1

ABSTRACT Naturally occurring polymorphisms in the protease of human immunodeficiency virus type 1 (HIV-1) subtype C would be expected to lead to adaptive (compensatory) changes in protease cleavage sites. To test this hypothesis, we examined the prevalences and patterns of cleavage site polymorphisms in the Gag, Gag-Pol, and Nef cleavage sites of C compared to those in non-C subtypes. Codon-based maximum-likelihood methods were used to assess the natural selection and evolutionary history of individual cleavage sites. Seven cleavage sites (p17/p24, p24/p2, NC/p1, NC/TFP, PR/RT, RT/p66, and p66/IN) were well conserved over time and in all HIV-1 subtypes. One site (p1/p6 gag ) exhibited moderate variation, and four sites (p2/NC, TFP/p6 pol , p6 pol /PR, and Nef) were highly variable, both within and between subtypes. Three of the variable sites are known to be major determinants of polyprotein processing and virion production. P2/NC controls the rate and order of cleavage, p6 gag is an important phosphoprotein required for virion release, and TFP/p6 pol , a novel cleavage site in the transframe domain, influences the specificity of Gag-Pol processing and the activation of protease. Overall, 58.3% of the 12 HIV-1 cleavage sites were significantly more diverse in C than in B viruses. When analyzed as a single concatenated fragment of 360 bp, 96.0% of group M cleavage site sequences fell into subtype-specific phylogenetic clusters, suggesting that they coevolved with the virus. Natural variation at C cleavage sites may play an important role, not only in regulation of the viral cycle but also in disease progression and response to therapy.

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