Biochemical and morphological characterization of the nuclear matrix from apoptotic HL-60 cells

Martelli, Alberto M.; Bortul, Roberta; Fackelmayer, Frank O.; Tazzari, Pier Luigi; Bareggi, Renato; Narducci, Paola; Zweyer, Marina

doi:10.1002/(sici)1097-4644(19990101)72:1<35::aid-jcb5>3.0.co;2-s

Cited by 23 publications

(24 citation statements)

References 60 publications

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“…IL-1␤-increased expression of lamin B and/or ribonucleoprotein K (ROK) (both identified from database match number 28) was NAME reversible in islets. Lamin B is primarily a structural nuclear-matrix protein that underlies the nuclear envelope; it has been studied in conjunction with induced apoptosis or necrosis (62), including that induced by 2D gel electrophoresis (63) in nonislet cells. ROK recruits molecular partners involved in transcriptional regulation and RNA processing (64).…”

Section: Discussionmentioning

confidence: 99%

Cytokine- or chemically derived nitric oxide alters the expression of proteins detected by two-dimensional gel electrophoresis in neonatal rat islets of Langerhans.

et al. 2000

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Interleukin-1␤ (IL-1␤) treatment of neonatal rat islets19 of a total of 1,600 detectable proteins in GSNOtreated islets (P < 0.01). We conclude 1) that the expression of up to 42 proteins is altered by cytokineinduced or chemically generated NO in the precise experimental conditions chosen and 2) that the majority of proteins altered by prior treatment with IL-1␤ may be the result of NO-independent IL-1␤-mediated regulation of gene expression. This study demonstrates that the combination of 2D gel electrophoresis and mass spectrometry is a powerful tool in the identification of ␤-cell proteins involved in the response to toxic mediators.

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Section: Discussionmentioning

confidence: 99%

Cytokine- or chemically derived nitric oxide alters the expression of proteins detected by two-dimensional gel electrophoresis in neonatal rat islets of Langerhans.

et al. 2000

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“…Proteolysis of lamins (29,30), NuMa (31)(32)(33), and SAF-A (10) appears to promote the destruction of the peripheral nuclear lamina as well as the internal nuclear scaffold that is thought to organize the genome. Investigation of these proteins sheds light on their role during apoptotic nuclear breakdown and, in addition, more clearly defines the function of these proteins in intact cells.…”

Section: Discussionmentioning

confidence: 99%

Apoptotic Cleavage of Scaffold Attachment Factor A (SAF-A) by Caspase-3 Occurs at a Noncanonical Cleavage Site

Kipp¹,

Schwab²,

Przybylski³

et al. 2000

Journal of Biological Chemistry

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Members of the caspase family of cysteine proteases play essential roles in the disintegration of cellular architecture during apoptosis. Caspases have been grouped into subfamilies according to their preferred cleavage sites, with the "apoptotic executioner" caspase-3 as the prototype of DEXD-dependent proteases. We show here that caspase-3 is more tolerant to variations of the cleavage site than previously anticipated and present an example of a noncanonical recognition site that is efficiently cleaved by caspase-3 in vitro and in vivo. The new cleavage site was identified in human scaffold attachment factor A, one of the major scaffold attachment region DNA-binding proteins of human cells thought to be involved in nuclear architecture by fastening chromatin loops to a proteinaceous nuclear skeleton, the so-called nuclear matrix or scaffold. Using an amino-terminal recombinant construct of scaffold attachment factor A and recombinant caspase-3, we have mapped the cleavage site by matrix-assisted laser desorption ionization/time of flight mass spectrometry and Edman sequencing. We find that cleavage occurs after Asp-100 in a sequence context (SALD) that does not conform to the hitherto accepted DEXD consensus sequence of caspase-3. A point mutation, D100A, abrogates cleavage by recombinant caspase-3 in vitro and during apoptosis in vivo, confirming SALD as a novel caspase-3 cleavage site.Apoptosis, or programmed cell death, is an active process of cellular self-destruction involved in normal development as well as in the maintenance of tissue homeostasis (reviewed in Ref. 1). Cells undergoing apoptosis show distinct morphological changes, including membrane blebbing, cytoplasmic and nuclear condensation, chromatin aggregation, and internucleosomal cleavage of DNA. In the final stages, the dying cells become fragmented into "apoptotic bodies," which are rapidly eliminated by phagocytic cells without eliciting significant inflammatory damage to surrounding cells. Many of the typical changes observed in cells undergoing apoptosis have been attributed to the limited proteolysis of a number of key substrates by a family of well conserved proteases, the caspases (reviewed in Refs. 2 and 3). Caspases are present in almost all eukaryotic cells as inactive zymogens that become activated after exposure to a variety of stimuli that lead to apoptosis. Activation occurs by one of two mechanisms, autocatalytic processing induced by their aggregation and/or cleavage by other caspases (2). Based on their substrate specificity, caspases have been grouped into three subfamilies (4). Group I caspases (e.g. caspase 1, 4, and 5) prefer the tetrapeptide sequence WEHD and have been implicated mainly in inflammatory processes rather than in apoptosis. Group II (e.g. caspases 2, 3, and 7) and group III (e.g. caspases 6, 8, 9, and 10) caspases have the recognition consensus sequence DEXD and (I/L/ V)EXD, respectively, and are involved in apoptosis. The different caspases have also been grouped by function into apoptotic initiators (casp...

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“…The first steps of this procedure are identical to that for nuclear matrix preparations (Zbarsky et al, 1962;Berezney et al, 1995). Therefore, IGC proteins were assumed to be components of the nuclear matrix and indeed, have been found in the nuclear matrix (Turner and Franchi, 1987;Jagatheesan et al, 1999;Martelli et al, 1999).…”

Section: Intranuclear Localisation Of P68mentioning

confidence: 99%

Satellite DNA binding and cellular localisation of RNA helicase P68

Enukashvily

Donev

Sheer

et al. 2005

Journal of Cell Science

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We purified a 68-kDa protein from the mouse nuclear matrix using ion exchange and affinity chromatography. Column fractions were tested for specific binding to mouse minor satellite DNA using a gel mobility shift assay. The protein was identified by mass spectrometry as RNA helicase P68. In fixed cells, P68 was found to shuttle in and out of SC35 domains, forming fibres and granules in a cell-cycle dependent manner. Analysis of the P68 sequence revealed a short potential coiled-coil domain that might be involved in the formation of P68 fibres. Contacts between centromeres and P68 granules were observed during all phases of the cycle but they were most prominent in mitosis. At this stage, P68 was found in both the centromeric regions and the connections between chromosomes. Direct interaction of P68/DEAD box RNA helicase with satellite DNAs in vitro has not been demonstrated for any other members of the RNA helicase family.

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Biochemical and morphological characterization of the nuclear matrix from apoptotic HL-60 cells

Cited by 23 publications

References 60 publications

Cytokine- or chemically derived nitric oxide alters the expression of proteins detected by two-dimensional gel electrophoresis in neonatal rat islets of Langerhans.

Cytokine- or chemically derived nitric oxide alters the expression of proteins detected by two-dimensional gel electrophoresis in neonatal rat islets of Langerhans.

Apoptotic Cleavage of Scaffold Attachment Factor A (SAF-A) by Caspase-3 Occurs at a Noncanonical Cleavage Site

Satellite DNA binding and cellular localisation of RNA helicase P68

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