Relationship between SUMO-1 modification of caspase-7 and its nuclear localization in human neuronal cells

Hayashi, Naoko; Shirakura, Hiromi; Uehara, Takashi; Nomura, Yasuyuki

doi:10.1016/j.neulet.2005.11.057

Cited by 32 publications

(26 citation statements)

References 29 publications

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“…One recent study found that sumoylation of a lysine in the N-terminal region of caspase-7, possibly within the KKKK motif, promotes nuclear localization (35). Our data support its function as a nuclear localization signal.…”

Section: Discussionsupporting

confidence: 84%

“…In Vitro Protein Synthesis and Cleavage-Caspase constructs were translated with 35 S-labeled methionine (Promega coupled kit) and incubated with 9.0 units of calpain-1 in Nonidet P40 buffer (19) with 5 M dithiothreitol (DTT) 2 and 20 mM CaCl 2 at 30°C for 30 min. Reactions were terminated by addition of EDTA, SDS sample buffer, and boiling.…”

Section: Methodsmentioning

confidence: 99%

“…Therefore, we tested whether calpains can directly cleave caspases and the impact of cleavage on caspase activity. In vitro-translated 35 S-labeled procaspases were treated with calpain-1 ( Fig. 1).…”

Section: Activity Of Caspase-7 Is Increased By Calpain-1 Cleavage-mentioning

confidence: 99%

“…Calpain-1 cleaves procaspases. In vitro-translated 35 S-labeled procaspases were treated with active calpain-1 to determine which caspases are calpain substrates. Each caspase, with and without calpain treatment, was run on a single gel and developed by autoradiography.…”

Section: Activity Of Caspase-7 Is Increased By Calpain-1 Cleavage-mentioning

confidence: 99%

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Calpain-1 Cleaves and Activates Caspase-7

Gafni

Cong

Chen

et al. 2009

Journal of Biological Chemistry

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Caspase-7 is an executioner caspase that plays a key role in apoptosis, cancer, and a number of neurodegenerative diseases. The mechanism of caspase-7 activation by granzyme B and caspase-3 has been well characterized. However, whether other proteases such as calpains activate or inactivate caspase-7 is not known. Here, we present that recombinant caspase-7 is directly cleaved by calpain-1 within the large subunit of caspase-7 to produce two novel products, large subunit p18 and p17. This new form of caspase-7 has a 6-fold increase in V max when compared with the previously characterized p20/p12 form. Zymography revealed that the smaller caspase-7 product (p17) is 18-fold more active than either the caspase-3-cleaved product (p20) or the larger calpain-1 product of caspase-7 (p18). Mass spectrometry and site-directed mutagenesis identified the calpain cleavage sites within the caspase-7 large subunit at amino acid 36 and 45/47. These proteolysis events occur in vivo as indicated by the accumulation of caspase-7 p18 and p17 subunits in cortical neurons undergoing Ca 2؉ dysregulation. Further, cleavage at amino acid 45/47 of caspase-7 by calpain results in a reduction in nuclear localization when compared with the caspase-3 cleavage product of caspase-7 (p20). Our studies suggest the calpain-activated form of caspase-7 has unique enzymatic activity, localization, and binding affinity when compared with the caspase-activated form.Apoptosis is a well-defined cellular destruction pathway that primarily utilizes a family of cysteine proteases, the caspases (1, 2). This cell death program can be initiated by cell death receptor activation (extrinsic pathway) or a variety of drugs or cellular stresses (intrinsic pathway) leading to activation of apical caspase-8, -9, and/or -10 (1, 3, 4). These initiator caspases in turn directly activate the executioner caspases, caspase-3 and -7, which through proteolysis of defined substrates are responsible for the dismantling of the cell and subsequent death (3, 4). Granzyme B, released by cytotoxic T lymphocytes to protect the host from pathogens and tumor cells, can also initiate this apoptotic cascade and therefore is considered an apical caspase mimic (5-7). All caspases, as well as granzyme B, preferentially cleave after aspartic acid residues, with many having well-defined consensus sequences, making substrate cleavage sites easy to predict and establish (3,4,7,8).Caspases exist in a latent form prior to activation. Both the initiator and executioner caspases are synthesized as a single chain protein, which require proteolytic cleavage to become active. Procaspase-7 is expressed as a 303-amino acid residue polypeptide chain. The activation and regulation of executioner caspase-7 by caspases and granzyme B has been extensively studied. Caspase-7 requires cleavage by caspase-3 and caspase-8/-10 or granzyme B, for activation (6, 9). Current evidence suggests that caspase-3 initially cleaves off the first 23 amino acids (propeptide, 2 kDa), followed by caspase-8/-10 or granzyme B...

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Section: Discussionsupporting

confidence: 84%

Section: Methodsmentioning

confidence: 99%

Section: Activity Of Caspase-7 Is Increased By Calpain-1 Cleavage-mentioning

confidence: 99%

Section: Activity Of Caspase-7 Is Increased By Calpain-1 Cleavage-mentioning

confidence: 99%

See 2 more Smart Citations

Calpain-1 Cleaves and Activates Caspase-7

Gafni

Cong

Chen

et al. 2009

Journal of Biological Chemistry

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“…This novel family of peptidases shares with caspases a common fold but displays radically different substrate specificity and it is not clear whether their members play an analogous role in programmed cell death in nonmetazoan organisms (66). Caspase-2 and -7 have been reported to be SUMOylated (67,68), and a detailed study on caspase-8 elegantly demonstrated that SUMOylation of this protein is determinant for its nuclear localization (69). Thus, modulation of metacaspase subcellular localization by SUMO conjugation is an intriguing possibility.…”

Section: Fig 5 Validation Of Metacaspase-3 As a Sumoylation Target mentioning

confidence: 99%

SUMOylation Pathway in Trypanosoma cruzi: Functional Characterization and Proteomic Analysis of Target Proteins

Bayona

Nakayasu

Laverrière

et al. 2011

Molecular & Cellular Proteomics

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SUMOylation is a relevant protein post-translational modification in eukaryotes. The C terminus of proteolytically activated small ubiquitin-like modifier (SUMO) is covalently linked to a lysine residue of the target protein by an isopeptide bond, through a mechanism that includes an E1-activating enzyme, an E2-conjugating enzyme, and transfer to the target, sometimes with the assistance of a ligase. The modification is reversed by a protease, also responsible for SUMO maturation. A number of proteins have been identified as SUMO targets, participating in the regulation of cell cycle progression, transcription, translation, ubiquitination, and DNA repair. In this study, we report that orthologous genes corresponding to the SUMOylation pathway are present in the etiological agent of Chagas disease, Trypanosoma cruzi. Furthermore, the SUMOylation system is functionally active in this protozoan parasite, having the requirements for SUMO maturation and conjugation. Immunofluorescence analysis showed that T. cruzi SUMO (TcSUMO) is predominantly found in the nucleus. To identify SUMOylation targets and get an insight into their physiological roles we generated transfectant T. cruzi epimastigote lines expressing a double-tagged T. cruzi SUMO, and SUMOylated proteins were enriched by tandem affinity chromatography. By two-dimensional liquid chromatography-mass spectrometry a total of 236 proteins with diverse biological functions were identified as potential T. cruzi SUMO targets. Of these, metacaspase-3 was biochemically validated as a bona fide SUMOylation substrate. Proteomic studies in other organisms have reported that orthologs of putative T.

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The Role of Protein SUMOylation in Neuronal Function

Wilkinson¹,

Henley

2010

Folding for the Synapse

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Relationship between SUMO-1 modification of caspase-7 and its nuclear localization in human neuronal cells

Cited by 32 publications

References 29 publications

Calpain-1 Cleaves and Activates Caspase-7

Calpain-1 Cleaves and Activates Caspase-7

SUMOylation Pathway in Trypanosoma cruzi: Functional Characterization and Proteomic Analysis of Target Proteins

The Role of Protein SUMOylation in Neuronal Function

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