Heat-Induced Changes in Nuclear-Associated Proteins in Normal and Thermotolerant HeLa Cells

Roti, Joseph L. Roti

doi:10.2307/3578735

Cited by 13 publications

(7 citation statements)

References 13 publications

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“…These thermolabile proteins (ie, those with T m 46ЊC-47ЊC) are distinct from the aggregated proteins associated with the nucleus as an insoluble fraction after heat shock (Roti Roti and Turkel 1994;Borrelli et al 1996). The results shown in Figure 12 for V79 cells and previously for mouse L cells (Borrelli et al 1996) indicate that the bulk of the protein associated with nuclei isolated from heatshocked cells is native.…”

Section: Discussionmentioning

confidence: 74%

“…Following heat shock, there is an increased binding of proteins to nuclear structures which was first detected as an increased protein/ DNA ratio of isolated nuclei (Roti Roti and Winward 1978;Tomasovic et al 1978). Although there is some specificity to the proteins found associated with the nucleus after heating, in general the amount of nearly all nuclear proteins is increased (Laszlo et al 1992; Roti Roti and Turkel 1994). This appears to be due to tighter binding of normally soluble or weakly bound nuclear proteins that ordinarily are lost from the nucleus of an unheated cell during isolation (Chu et al 1993;Borrelli et al 1996).…”

Section: Introductionmentioning

confidence: 99%

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The nuclear matrix is a thermolabile cellular structure

Lepock

Frey²,

Heynen³

et al. 2001

Cell Stress Chaper

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Heat shock sensitizes cells to ionizing radiation, cells heated in S phase have increased chromosomal aberrations, and both Hsp27 and Hsp70 translocate to the nucleus following heat shock, suggesting that the nucleus is a site of thermal damage. We show that the nuclear matrix is the most thermolabile nuclear component. The thermal denaturation profile of the nuclear matrix of Chinese hamster lung V79 cells, determined by differential scanning calorimetry (DSC), has at least 2 transitions at Tm = 48 degrees C and 55 degrees C with an onset temperature of approximately 40 degrees C. The heat absorbed during these transitions is 1.5 cal/g protein, which is in the range of enthalpies for protein denaturation. There is a sharp increase in 1-anilinonapthalene-8-sulfonic acid (ANS) fluorescence with Tm = 48 degrees C, indicating increased exposure of hydrophobic residues at this transition. The Tm = 48 degrees C transition has a similar Tm to those predicted for the critical targets for heat-induced clonogenic killing (Tm = 46 degrees C) and thermal radiosensitization (Tm = 47 degrees C), suggesting that denaturation of nuclear matrix proteins with Tm = 48 degrees C contribute to these forms of nuclear damage. Following heating at 43 degrees C for 2 hours, Hsc70 binds to isolated nuclear matrices and isolated nuclei, probably because of the increased exposure of hydrophobic domains. In addition, approximately 25% of exogenous citrate synthase also binds, indicating a general increase in aggregation of proteins onto the nuclear matrix. We propose that this is the mechanism for increased association of nuclear proteins with the nuclear matrix observed in nuclei Isolated from heat-shocked cells and is a form of indirect thermal damage.

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Section: Discussionmentioning

confidence: 74%

Section: Introductionmentioning

confidence: 99%

The nuclear matrix is a thermolabile cellular structure

Lepock

Frey²,

Heynen³

et al. 2001

Cell Stress Chaper

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“…The translocation of proteins from the cytoplasm to the nuclear compartment in response to heat shock has been well documented [Roti Roti and Turkel, 1994;VanderWaal et al, 1996]. The results shown in Figures 1 and 2 suggested that cells subjected to heat shock demonstrated elevated CK2 in the nuclear matrix isolated from these cells, and this response appeared to be due to shuttling of the kinase from the cytoplasm to the nuclear matrix in response to heat shock.…”

Section: Evidence For Intranuclear Trafficking Of Ck2 In Response To mentioning

confidence: 76%

Heat shock mediated modulation of protein kinase CK2 in the nuclear matrix*

Davis

Wang

Zhang

et al. 2002

J of Cellular Biochemistry

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Nuclear matrix, a key structure in the nuclear framework, appears to be a particularly responsive target during heat shock treatment of cells. We have previously shown that nuclear matrix is a preferential target for protein kinase CK2 signaling in the nucleus. The levels of CK2 in the nuclear matrix undergo dynamic changes in response to altered growth status in the cell. Here, we have demonstrated that CK2 targeting to the nuclear matrix is profoundly influenced by treatment of the cells to temperatures higher than 37 degrees C. Rapid increase in the nuclear matrix association of CK2 is observed when cells are placed at temperatures of 41 and 45 degrees C. This effect at 45 degrees C was higher than at 41 degrees C, and was time-dependent. Also, different cell lines behaved in a qualitatively similar manner though the quantitative responses differed. The modulations in the nuclear matrix associated CK2 in response to heat shock appear to be due to trafficking of the enzyme between cytosolic and nuclear compartments. In addition, it was noted that isolated nuclei subjected to heat shock also responded by a shuttling of the intrinsic CK2 to the nuclear matrix compartment. These results suggest that modulations in CK2 in the nuclear compartment in response to the heat stress occur not only by a translocation of the enzyme from the cytoplasmic compartment to the nuclear compartment, but also that there is a redistribution of the kinase within the nuclear compartment resulting in a preferential association with the nuclear matrix. The results support the notion that CK2 association with the nuclear matrix in response to heat shock may serve a protective role in the cell response to stress.

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“…Several mitogenic signal or cell cycle control proteins appear to be kept in a certain conformation by such multiprotein complexes [36]. Chaperones of the HSP70 family also help proteins to enter the nucleus and to associate with or dissociate from other nuclear protein molecules [16]. Furthermore, this chaperone family is synthesized in a cell-cycle-dependent manner with highest concentrations within S phase [42][43][44][45].…”

Section: Heat Shock Responsementioning

confidence: 99%

Heat shock effects on cell cycle progression

Kühl¹,

Rensing²

2000

CMLS, Cell. Mol. Life Sci.

120

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In mammalian cells, short-term (acute) exposure to a moderate heat shock leads to a transient arrest of cells at mainly two cell cycle checkpoints, the G1/S and G2/M transitions. This is documented by the more or less synchronous resumption of cell cycle progression from these checkpoints during recovery. The reason for the accumulation of cells at these checkpoints may be found in activity thresholds of cyclin-dependent kinases (Cdks) at both transitions which are determined by (i) the amounts of the responsible cyclins, (ii) regulatory phosphorylation of the Cdks and (iii) the inhibition of Cdks by associated regulatory proteins (Ckis). All three regulatory systems may be subject to heat-shock-dependent changes, the amounts of Ckis, in particular, being increased. Cdk-dependent phosphorylation of the retinoblastoma protein and the subsequent release of active S-phase-specific transcription factors E2F/DP are considered as major heat-sensitive steps in cell cycle progression. Furthermore, high acute heat shock and long-term (chronic) heat treatment may lead to cell-type-specific forms of cell death. All types of responses to heat treatment are subject to adaptation after a 'priming' treatment, probably due to higher levels of heat shock proteins.

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Heat-Induced Changes in Nuclear-Associated Proteins in Normal and Thermotolerant HeLa Cells

Cited by 13 publications

References 13 publications

The nuclear matrix is a thermolabile cellular structure

The nuclear matrix is a thermolabile cellular structure

Heat shock mediated modulation of protein kinase CK2 in the nuclear matrix*

Heat shock effects on cell cycle progression

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