A highly conserved family of chromosomal proteins are caspase substrates during apoptosis

Research output: Contribution to journalArticle

Abstract

Apoptosis, or programmed cell death (PCD), is an essential component of embryogenesis, immune system function, and tissue homeostasis in eukaryotic organisms. This process is mediated by, and absolutely requires, the activity of caspases, cysteine endoproteases that recognize specific tetrapeptide sequences having an aspartate residue at the P1 location of the cleavage site. A morphological hallmark of this process is the appearance of condensed and fragmented nuclei. However, the mechanism bridging caspase activation with apoptotic chromatin condensation is unknown. In this report, we demonstrate that FIN16, a mouse protein encoded by the fibroblast growth factor-inducible cDNA 16 (FIN16), is a novel caspase substrate. A BLAST search of the nonredundant GenBank database and subsequent sequence alignment revealed high homology between FIN16 and the carboxy terminal domains of structural maintenance of chromosomes (SMC) family proteins, including the Saccharomyces cerevisiae SMC2, Xenopus laevis XCAP-E condensin, and Gallus gallus Scll chromosomal scaffold proteins. Importantly, all of these proteins contain a potential caspase-3 cleavage sequence, DEVD. In vitro, purified caspase-3 cleaved radiolabelled FIN16 generating two fragments, one of which was probably generated by cleavage after D305, as expected. However, FIN16 did not serve as a substrate for caspase-8, but was cleaved by granzyme B at a different site than D305. Furthermore, apoptotic cytosolic extracts from Fas/CD95- and staurosporine-activated cells were also able to process the FIN16 protein. This was fully inhibited by the tetrapeptide inhibitors Z-VAD-FMK and Z-DEVD-FMK, confirming the involvement of caspases. The presence of an evolutionarily conserved caspase consensus sequence DEV/ID in SMC proteins further implicates the caspase-mediated cleavage of these proteins as a potential mechanism of apoptotic chromatin condensation and/or destruction of chromosomal structure.

Original languageEnglish (US)
JournalJournal of Investigative Medicine
Volume47
Issue number2
StatePublished - Feb 1999

Fingerprint

Caspases
Complementary DNA
Apoptosis
Substrates
Proteins
Caspase 3
Chromatin
Chromosomes
Tissue homeostasis
Maintenance
Condensation
Saccharomyces cerevisiae Proteins
Granzymes
Staurosporine
Fibroblast Growth Factors
Caspase 8
Sequence Alignment
Nucleic Acid Databases
Consensus Sequence
Xenopus laevis

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

@article{dd83fc77a00f4006a3d9f8263ae49e6c,
title = "A highly conserved family of chromosomal proteins are caspase substrates during apoptosis",
abstract = "Apoptosis, or programmed cell death (PCD), is an essential component of embryogenesis, immune system function, and tissue homeostasis in eukaryotic organisms. This process is mediated by, and absolutely requires, the activity of caspases, cysteine endoproteases that recognize specific tetrapeptide sequences having an aspartate residue at the P1 location of the cleavage site. A morphological hallmark of this process is the appearance of condensed and fragmented nuclei. However, the mechanism bridging caspase activation with apoptotic chromatin condensation is unknown. In this report, we demonstrate that FIN16, a mouse protein encoded by the fibroblast growth factor-inducible cDNA 16 (FIN16), is a novel caspase substrate. A BLAST search of the nonredundant GenBank database and subsequent sequence alignment revealed high homology between FIN16 and the carboxy terminal domains of structural maintenance of chromosomes (SMC) family proteins, including the Saccharomyces cerevisiae SMC2, Xenopus laevis XCAP-E condensin, and Gallus gallus Scll chromosomal scaffold proteins. Importantly, all of these proteins contain a potential caspase-3 cleavage sequence, DEVD. In vitro, purified caspase-3 cleaved radiolabelled FIN16 generating two fragments, one of which was probably generated by cleavage after D305, as expected. However, FIN16 did not serve as a substrate for caspase-8, but was cleaved by granzyme B at a different site than D305. Furthermore, apoptotic cytosolic extracts from Fas/CD95- and staurosporine-activated cells were also able to process the FIN16 protein. This was fully inhibited by the tetrapeptide inhibitors Z-VAD-FMK and Z-DEVD-FMK, confirming the involvement of caspases. The presence of an evolutionarily conserved caspase consensus sequence DEV/ID in SMC proteins further implicates the caspase-mediated cleavage of these proteins as a potential mechanism of apoptotic chromatin condensation and/or destruction of chromosomal structure.",
author = "Tepper, {Clifford G} and Seldin, {Michael F}",
year = "1999",
month = "2",
language = "English (US)",
volume = "47",
journal = "Journal of Investigative Medicine",
issn = "1081-5589",
publisher = "Lippincott Williams and Wilkins",
number = "2",

}

TY - JOUR

T1 - A highly conserved family of chromosomal proteins are caspase substrates during apoptosis

AU - Tepper, Clifford G

AU - Seldin, Michael F

PY - 1999/2

Y1 - 1999/2

N2 - Apoptosis, or programmed cell death (PCD), is an essential component of embryogenesis, immune system function, and tissue homeostasis in eukaryotic organisms. This process is mediated by, and absolutely requires, the activity of caspases, cysteine endoproteases that recognize specific tetrapeptide sequences having an aspartate residue at the P1 location of the cleavage site. A morphological hallmark of this process is the appearance of condensed and fragmented nuclei. However, the mechanism bridging caspase activation with apoptotic chromatin condensation is unknown. In this report, we demonstrate that FIN16, a mouse protein encoded by the fibroblast growth factor-inducible cDNA 16 (FIN16), is a novel caspase substrate. A BLAST search of the nonredundant GenBank database and subsequent sequence alignment revealed high homology between FIN16 and the carboxy terminal domains of structural maintenance of chromosomes (SMC) family proteins, including the Saccharomyces cerevisiae SMC2, Xenopus laevis XCAP-E condensin, and Gallus gallus Scll chromosomal scaffold proteins. Importantly, all of these proteins contain a potential caspase-3 cleavage sequence, DEVD. In vitro, purified caspase-3 cleaved radiolabelled FIN16 generating two fragments, one of which was probably generated by cleavage after D305, as expected. However, FIN16 did not serve as a substrate for caspase-8, but was cleaved by granzyme B at a different site than D305. Furthermore, apoptotic cytosolic extracts from Fas/CD95- and staurosporine-activated cells were also able to process the FIN16 protein. This was fully inhibited by the tetrapeptide inhibitors Z-VAD-FMK and Z-DEVD-FMK, confirming the involvement of caspases. The presence of an evolutionarily conserved caspase consensus sequence DEV/ID in SMC proteins further implicates the caspase-mediated cleavage of these proteins as a potential mechanism of apoptotic chromatin condensation and/or destruction of chromosomal structure.

AB - Apoptosis, or programmed cell death (PCD), is an essential component of embryogenesis, immune system function, and tissue homeostasis in eukaryotic organisms. This process is mediated by, and absolutely requires, the activity of caspases, cysteine endoproteases that recognize specific tetrapeptide sequences having an aspartate residue at the P1 location of the cleavage site. A morphological hallmark of this process is the appearance of condensed and fragmented nuclei. However, the mechanism bridging caspase activation with apoptotic chromatin condensation is unknown. In this report, we demonstrate that FIN16, a mouse protein encoded by the fibroblast growth factor-inducible cDNA 16 (FIN16), is a novel caspase substrate. A BLAST search of the nonredundant GenBank database and subsequent sequence alignment revealed high homology between FIN16 and the carboxy terminal domains of structural maintenance of chromosomes (SMC) family proteins, including the Saccharomyces cerevisiae SMC2, Xenopus laevis XCAP-E condensin, and Gallus gallus Scll chromosomal scaffold proteins. Importantly, all of these proteins contain a potential caspase-3 cleavage sequence, DEVD. In vitro, purified caspase-3 cleaved radiolabelled FIN16 generating two fragments, one of which was probably generated by cleavage after D305, as expected. However, FIN16 did not serve as a substrate for caspase-8, but was cleaved by granzyme B at a different site than D305. Furthermore, apoptotic cytosolic extracts from Fas/CD95- and staurosporine-activated cells were also able to process the FIN16 protein. This was fully inhibited by the tetrapeptide inhibitors Z-VAD-FMK and Z-DEVD-FMK, confirming the involvement of caspases. The presence of an evolutionarily conserved caspase consensus sequence DEV/ID in SMC proteins further implicates the caspase-mediated cleavage of these proteins as a potential mechanism of apoptotic chromatin condensation and/or destruction of chromosomal structure.

UR - http://www.scopus.com/inward/record.url?scp=33750139717&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33750139717&partnerID=8YFLogxK

M3 - Article

AN - SCOPUS:33750139717

VL - 47

JO - Journal of Investigative Medicine

JF - Journal of Investigative Medicine

SN - 1081-5589

IS - 2

ER -