Regulation of nuclear scaffold protease activity by calcium

Madsen, Kenneth R.; Fiskum, Gary; Clawson, Gary A.

doi:10.1016/0014-4827(90)90103-h

Cited by 17 publications

(10 citation statements)

References 18 publications

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“…The major lamin-hydrolyzing proteolytic activity in rat nuclei requires Ca, is stimulated by micromolar Ca, and is inhibited by calmodulin antagonists (Madsen et al, 1990). Whether the rat NTPase itself, like the pea NTPase, can be stimulated by calmodulin has not been determined, nor is it known whether the protease activity found in pea nuclei can be activated by Ca or calmodulin.…”

Section: Discussionmentioning

confidence: 99%

The Major Nucleoside Triphosphatase in Pea (Pisum sativum L.) Nuclei and in Rat Liver Nuclei Share Common Epitopes Also Present in Nuclear Lamins

Tong

Dauwalder

Clawson³

et al. 1993

Plant Physiol.

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l h e major nucleoside triphosphatase (NTPase) activities in mammalian and pea (Pisum sativum 1.) nuclei are associated with enzymes that are very similar both biochemically and immunochemically. l h e major NTPase from rat liver nuclei appears to be a 46-kD enzyme that represents the N-terminal portion of lamins A and C, two lamina proteins that apparently arise from the same gene by alternate splicing. Monoclonal antibody (MAb) C2, raised to human lamin C, both immunoprecipitates the major (47 kD) NTPase in pea nuclei and recognizes i t in western blot analyses. A polyclonal antibody preparation raised to the 47-kD pea NTPase (pc480) reacts with the same lamin bands that are recognized by MAb C2 in mammalian nuclei. l h e pc480 antibodies also bind to the same lamin-like bands in pea nuclear envelope-matrix preparations that are recognized by C2 and three other MAbs known to bind to mammalian lamins. In immunofluorescence assays, pc480 and anti-lamin antibodies stain both cytoplasmic and nuclear antigens in plant cells, with slightly enhanced staining along the periphery of the nuclei. These results indicate that the pea and rat liver NTPases are structurally similar and that, in pea nuclei as in rat liver nuclei, the major NTPase i s probably derived from a lamin precursor by proteolysis.

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Section: Discussionmentioning

confidence: 99%

The Major Nucleoside Triphosphatase in Pea (Pisum sativum L.) Nuclei and in Rat Liver Nuclei Share Common Epitopes Also Present in Nuclear Lamins

Tong

Dauwalder

Clawson³

et al. 1993

Plant Physiol.

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“…Previous work has shown that calmodulin antagonists are capable of inhibiting DNA fragmentation in thymocytes exposed to apoptotic triggers (29,39), and it has been suggested that the NS protease may be regulated by calmodulin (40). We therefore tested the effects of the calmodulin antagonist calmidizolium on DNA fragmentation and lamin B 1 cleavage in isolated nuclei.…”

Section: Nuclear Scaffold Protease In Ca 2ϩ -Mediated Endonuclease Acmentioning

confidence: 99%

Calcium-dependent, Interleukin 1β-converting Enzyme Inhibitor-insensitive Degradation of Lamin B1 and DNA Fragmentation in Isolated Thymocyte Nuclei

McConkey¹

1996

Journal of Biological Chemistry

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Recent work suggests that the proteolytic degradation of the nuclear lamins is a common event in apoptosis, although the nature of the proteases involved is still not clear. Our previous work showed that the degradation of lamin B 1 in glucocorticoid-treated thymocytes occurs via a Ca 2؉ -sensitive mechanism and that exogenous Ca 2؉ promotes lamin degradation in isolated thymocyte nuclei from untreated cells. Here we demonstrate that peptide-based inhibitors of the interleukin 1␤-converting enzyme family of cysteine proteases (TyrVal-Ala-Asp fluoromethyl ketone) and of the nuclear scaffold multicatalytic proteinase (Ala-Pro-Phe chloromethyl ketone) block the degradation of lamin B 1 to a 21-kDa fragment in thymocytes treated with glucocorticoid, the Ca 2؉ -mobilizing agent thapsigargin, or antibodies to the T cell receptor. However, among a panel of inhibitors specific for several different proteases implicated in apoptosis, only tosylphenylalanyl chloromethyl ketone and the nuclear scaffold protease inhibitor block lamin degradation, histone H1 cleavage, and DNA fragmentation in isolated thymocyte nuclei incubated with Ca 2؉ . Overexpression of human BCL-2 in nuclei by stable transfection resulted in an inhibition of Ca 2؉ -stimulated lamin degradation and DNA fragmentation, suggesting that endogenous nuclear BCL-2 regulates activation of the nuclear scaffold protease. The results demonstrate the existence of an alternative pathway of lamin degradation and DNA fragmentation mediated by a resident Ca 2؉ -stimulated nuclear protease that is not directly dependent upon activation of the interleukin 1␤-converting enzyme family of cell death regulators.Apoptosis is a process of physiological cell death characterized by a series of stereotyped morphological alterations and endogenous endonuclease activation leading to the generation of large (50 kb 1 ) and oligonucleosomal DNA fragments (1, 2). Although inhibitors of macromolecular synthesis prevent apoptosis in many circumstances, it is currently thought that the essential enzymatic components of the effector pathway are constitutively expressed in all mammalian cells beyond the blastomere stage of embryonic development (3). The identities of these cell death mediators are currently not known, but a major recent breakthrough toward their identification has come from work on the genes that regulate programmed cell death in the nematode Caenorhabditis elegans (4). This work has revealed that the C. elegans death promoter ced-3 is structurally homologous to a growing family of proteins related to human interleukin 1␤-converting enzyme (ICE) (5), a cysteine protease. Subsequent research has shown that specific viral and peptide-based inhibitors of the ICE family block apoptotic cell death in many mammalian models (4 -9), indicating that ICE and its homologs play crucial roles in the effector phase of apoptosis.Recent work has shown that the nuclear lamins are cleaved by one or more members of the ICE family during apoptosis. Early work by Kaufmann (10) demonstrated th...

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“…RNA was purified from the supernate using guanidine isothiocyanate and CsCl (19) and resuspended and reprecipitated twice with 70% ethanol after addition of sodium acetate to 0.3 mol/L; RNA was quantitated by absorbance at 260 nm, and preparations had &60,280 ratios of 2. NS was prepared from purified nuclear pellets in the presence of protease inhibitors (17,18), except for the final rinse. Protein was quantitated by the method of Lowry et al (20), and NS preparations were examined by standard SDS-PAGE (21).…”

Section: Methodsmentioning

confidence: 99%

Alterations in nuclear scaffold constituents during carbon tetrachloride-induced liver regeneration

et al. 1991

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Liver regeneration was induced in rats by treatment with CCl4, which results in substantial regenerative activity with a sharp mitotic response 2 days after intoxication. Closely paralleling the mitotic index, we observed fourfold increases in nuclear scaffold nucleoside triphosphatase, an activity thought to participate in nucleocytoplasmic RNA transport and in the 46 kD putative enzyme and its selective photolabeling. Because previous work has indicated that the 46 kD protein may be proteolytically derived from lamins A/C by cleavage at a tyrosine residue at aa376, we investigated the response of lamin A/C transcripts during this regeneration. Surprisingly, Northern blot analyses after CCl4 administration showed low levels of lamin A/C transcripts (which appeared to be predominantly poly[A]-), and we found a decrease in immunoprecipitable lamins A/C from in vitro translation of poly(A)- selected RNA. To circumvent potential problems with such analyses, we used reverse transcription/polymerase chain reaction amplification of lamin A/C transcripts from total cytoplasmic RNA. These assays showed a transient, comparatively minor increase in lamin A/C transcripts 1 day after treatment, but levels rapidly declined from 1 to 3 days and were decreased at 3 to 5 days. However, nuclear scaffold protease activity, which shows a considerable selectivity for lamins A/C and may be involved in derivation of the 46 kD protein, increased in parallel to the mitotic response and increases in nucleoside triphosphatase, as assessed using a nonspecific (Azocoll) protease assay. Assays with a specific tyrosine-containing substrate (Z-Y-Sbenzyl) showed an increase that mirrored that observed with the nonspecific substrate.(ABSTRACT TRUNCATED AT 250 WORDS)

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Regulation of nuclear scaffold protease activity by calcium

Cited by 17 publications

References 18 publications

The Major Nucleoside Triphosphatase in Pea (Pisum sativum L.) Nuclei and in Rat Liver Nuclei Share Common Epitopes Also Present in Nuclear Lamins

The Major Nucleoside Triphosphatase in Pea (Pisum sativum L.) Nuclei and in Rat Liver Nuclei Share Common Epitopes Also Present in Nuclear Lamins

Calcium-dependent, Interleukin 1β-converting Enzyme Inhibitor-insensitive Degradation of Lamin B1 and DNA Fragmentation in Isolated Thymocyte Nuclei

Alterations in nuclear scaffold constituents during carbon tetrachloride-induced liver regeneration

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