A nuclear protein associated with lethal heat shock of HL-60 cells

Pipkin, James L.; Hinson, William G.; Lyn‐Cook, Lascelles E.; Burns, E. Robert; Sheehan, Daniel M.; Casciano, Daniel A.

doi:10.1016/0014-4827(92)90416-6

Cited by 8 publications

(10 citation statements)

References 30 publications

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“…2. The unit was fabricated in-house and was used in previous research [13]. The electrode plates (upper and lower units) were composed of graphite impregnated electrical conducting plastic.…”

Section: V8 Protease Digestsmentioning

confidence: 99%

“…The upper plate was connected to the cathode (-) and the lower plate to the anode (+) source. All proteins in this buffer system run as anode particles [13,36,371. The nitrocellulose membrane was cut to the exact size of the microslab gel (25 X 35 mm) and the filter and spacer sheets were slightly smaller (23 X 33 mm).…”

Section: V8 Protease Digestsmentioning

confidence: 99%

“…Ultramicro-electrophoresis (UMEA) was developed by us and has been used in various applications including the studies of nuclear protein metabolism in embryonic murine limb bud development 17, 81, and in the differentiation of HL60 cells 19, 101 following administration of retinoic acid (RA), a vitamin A analog [ll]. Most recently we have used UMEA for the PAGE analysis of microsamples from nuclei sorted from partitions of the cell cycle [9,10,[12][13][14][15]. Microelectrophoresis has facilitated the investigation of nuclear protein synthesis and phosphorylation from partitions of the cell cycle [9,10,13,14,[16][17][18], and has validated the utility of these processes as biomarkers for elucidating the alteration of cellular metabolism induced by chemicals/toxic stress.…”

Section: Introductionmentioning

confidence: 99%

“…Most recently we have used UMEA for the PAGE analysis of microsamples from nuclei sorted from partitions of the cell cycle [9,10,[12][13][14][15]. Microelectrophoresis has facilitated the investigation of nuclear protein synthesis and phosphorylation from partitions of the cell cycle [9,10,13,14,[16][17][18], and has validated the utility of these processes as biomarkers for elucidating the alteration of cellular metabolism induced by chemicals/toxic stress. We have fabricated an ultramicro trans-blot cell, described in detail for the first time in this paper, which was employed by us in immunological recognition of antigens separated in the second-dimensional gels of UMEA [13].…”

Section: Introductionmentioning

confidence: 99%

“…Microelectrophoresis has facilitated the investigation of nuclear protein synthesis and phosphorylation from partitions of the cell cycle [9,10,13,14,[16][17][18], and has validated the utility of these processes as biomarkers for elucidating the alteration of cellular metabolism induced by chemicals/toxic stress. We have fabricated an ultramicro trans-blot cell, described in detail for the first time in this paper, which was employed by us in immunological recognition of antigens separated in the second-dimensional gels of UMEA [13]. The purpose of this study was twofold: first, to study the preliminary biochemical characteristics and possible homology of a unique nuclear protein (M, 34 000) expressed in the hypothalamus of old caloric restricted rats (O/CR) and in embryonic mouse brain elicited by diverse stressors; second, to describe the features and operation of the transfer cell used in immunochemical analysis in this and previous studies.…”

Section: Introductionmentioning

confidence: 99%

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The homology of a novel polypeptide with stress protein characteristics in embryonic mice brain and in the hypothalamus of caloric restricted rats as determined by ultramicro western blotting

et al. 1994

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A novel protein (p34) was observed in polyacrylamide gel fluorographs of gestation day 13 embryonic mouse brain following retinoic acid dosing of dams. Another p34 polypeptide with identical gel migratory characteristics was seen in the hypothalamus of old caloric restricted rats after "food deprivation stress". Western blotting, employing an ultramicro trans-blot cell developed in our laboratory, detected identical immunochemical determinants between these proteins, verifying their homology. Peptide mapping and Western blotting further validated the uniqueness of p34 compared with other stress proteins including heme oxygenase.

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Section: V8 Protease Digestsmentioning

confidence: 99%

Section: V8 Protease Digestsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

The homology of a novel polypeptide with stress protein characteristics in embryonic mice brain and in the hypothalamus of caloric restricted rats as determined by ultramicro western blotting

et al. 1994

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Characterization of a new high-temperature-induced 66-kDa heat-shock protein, antigenically related to heat-shock protein 72

Delpino¹,

Mileo²,

Lapenta³

et al. 1996

J. Cell. Biochem.

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M-14 human melanoma cells, following severe hyperthermic exposures, synthesized a heat-shock protein of 66 kDa (hsp 66), in addition to the major "classic" heat-shock proteins. This hsp 66 was not expressed following mild hyperthermic exposures sufficient to trigger the synthesis of the other heat-shock proteins. The induction of hsp 66 was observed also in Li human glioma cells treated at 45 degrees C for 20 min. By contrast, hsp 66 was not induced in seven other human cell lines (both melanoma and nonmelanoma) when they were subjected to the same hyperthermic treatment. Immunological recognition experiments showed that hsp 66 cross-reacted with the inducible hsp 72, but not with the constitutive hsp 73. The possibility that hsp 66 is a breakdown product of hsp 72 was ruled out by the fact that Poly(A)+ RNA extracted from cells treated at 45 degrees C for 20 min was able to direct the synthesis of hsp 66 (together with hsp 72) in a message-dependent rabbit reticulocyte lysate, as well as in microinjected Xenopus oocytes. By contrast, only the hsp 72 was expressed using Poly(A)+ RNA extracted from cells heated at 42 degrees C for 1 h. Affinity chromatography experiments on ATP-agarose showed that hsp 66 did not bind ATP in vitro. hsp 66 was localized both in the cytoplasm (cytosol, mitochondria, and microsome fraction) and in the nuclei of cells recovered from a severe heat shock: this intracellular distribution closely corresponded to that of hsp 72. The nuclear-associated hsp 66 was found to be tightly bound to nuclear structures and could not be extracted by incubation in ATP-containing buffer.

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Induction of stress proteins by electromagnetic fields in cultured HL-60 cells

Pipkin

Hinson

Young

et al. 1999

Bioelectromagnetics

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HL‐60 cells in culture were exposed for 2 h to a sinusoidal 0.1 or 1 mT (1 or 10 Gauss) magnetic field at 60 Hz and pulse labeled after exposure with radioactive isotopes by incubation by using either [35S]methionine, [3H]leucine, or [33P]phosphate. The radioactive labels were incorporated into cellular proteins through synthesis or phosphorylation. Proteins were extracted from electrostatically sorted nuclei, and the heat shock/stress proteins (sp) were analyzed for synthesis and phosphorylation by two‐dimensional polyacrylamide gel electrophoresis. In the control cultures (no exposure to the magnetic field), sp 72c (cognate form) was faintly observed. A 0.1 mT exposure did not show sp metabolism to be different from that of the controls; however, after a 1 mT exposure of the HL‐60 cells, sp 70i (inducible form) was synthesized ([35S]methionine incorporation). Sp 90 was not synthesized at either field level, but was phosphorylated ([33P]phosphate incorporation) in the 1 mT exposure. Sp 27 (isoforms a and b) was induced after a 1 mT exposure as reflected by labeling with [3H]leucine. These sps were not detected after a 0.1 mT exposure. After a 1 mT exposure and labeling with [33P], sp 27 isoforms b and c were phosphorylated whereas isoform ‘a‘ was not observed. Sps 70i, 72c, and 90 were identified by commercial sp antibodies. Likewise, polypeptides a, b, and c were verified as sp 27 isoforms by Western blotting. Statistical evaluation of sp areas and densities, determined from fluorographs by Western‐blot analysis, revealed a significant increase in sps 90 and 27a after a 1 mT magnetic field exposure. The 1 mT magnetic field interacts at the cellular level to induce a variety of sp species. Bioelectromagnetics 20:347–357, 1999. Published 1999 Wiley‐Liss, Inc.

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A nuclear protein associated with lethal heat shock of HL-60 cells

Cited by 8 publications

References 30 publications

The homology of a novel polypeptide with stress protein characteristics in embryonic mice brain and in the hypothalamus of caloric restricted rats as determined by ultramicro western blotting

The homology of a novel polypeptide with stress protein characteristics in embryonic mice brain and in the hypothalamus of caloric restricted rats as determined by ultramicro western blotting

Characterization of a new high-temperature-induced 66-kDa heat-shock protein, antigenically related to heat-shock protein 72

Induction of stress proteins by electromagnetic fields in cultured HL-60 cells

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