We have examined and compared a number of cellular and biochemical events associated with the recovery process of rat fibroblasts placed under stress by different agents. Metabolic pulse-labeling studies of cells recovering from either heat-shock treatment, exposure to sodium arsenite, or exposure to an amino acid analogue of proline, L-azetidine 2-carboxylic acid, revealed interesting differences with respect to the individual stress proteins produced, their kinetics of induction, as well as the decay in their synthesis during the recovery period. In the initial periods of recovery, the major stress-induced 72-kD protein accumulates within the altered nucleoli in close association with the pre-ribosomal-containing granular region. During the later times of recovery from stress, the nucleoli begin to regain a normal morphology, show a corresponding loss of the 72-kD protein, and the majority of the protein now begins to accumulate within the cytoplasm in three distinct locales: the perinuclear region, along the perimeter of the cells, and finally in association with large phase-dense structures. These latter structures appear to consist of large aggregates of phase-dense material with no obvious encapsulating membrane. More interestingly we show, using double-label indirect immunofluorescence analysis, that much of the perinuclear and cell perimeter-distributed 72-kD protein coincides with the distribution of the cytoplasmic ribosomes. We discuss the possible implications of the presence of the 72-kD stress proteins within the pre-ribosomal-containing granular region of the nucleolus as well as its subsequent colocalization with cytoplasmic ribosomes in terms of the translational changes which occur in cells both during and after recovery from physiological stress.

/pdf/cellular-and-biochemical-events-in-mammalian-cells-during-3s5i8ja8st.pdf

Cellular and biochemical events in mammalian cells during and after recovery from physiological stress.

Living organisms are known to react to a heat stress by the selective induction in the synthesis of several polypeptides. In this review we list the major stress proteins of mammalian cells that are induced by heat shock and other environments and categorize these proteins into specific subgroups: the major heat shock proteins, the glucose-regulated proteins, and the low-molecular-weight heat shock proteins. Characteristics of the localization and expression of proteins in each of these subgroups are presented. Specifically, the nuclear/nucleolar locale of certain of the major heat shock proteins is considered with respect to their association with RNA and the recovery of cells after a heat exposure. The induction of these major heat shock proteins and the repression of the glucose-regulated proteins as a result of reoxygenation of anoxic cells or by the addition of glucose to glucose-deprived cultures is described. Changes in the expression of these protein systems during embryogenesis and differentiation in mammalian and nonmammalian systems is summarized, and the protective role that some of these proteins appear to play in protecting the animal against the lethal effects of a severe heat treatment and against teratogenesis is critically examined.

https://www.physiology.org/doi/pdf/10.1152/ajpcell.1986.250.1.C1

Stress protein systems of mammalian cells.

Nuclear and nucleolar localization of the 72,000-dalton heat shock protein in heat-shocked mammalian cells.

Using both electron microscopy and immunological methods, we have characterized a number of changes occurring in rat fibroblasts after heat-shock treatment. Incubation of the cells for 3 h at 42 degrees-43 degrees C resulted in a number of changes within the cytoplasm including: a disruption and fragmentation of the Golgi complex; a modest swelling of the mitochondria and subtle alterations in the packing of the cristae; and alterations in cytoskeletal elements, specifically a collapse and aggregation of the vimentin-containing intermediate filaments around the nucleus. A number of striking changes were also found within the nuclei of the heat-treated cells: (a) We observed the appearance of rod-shaped bodies consisting of densely packed filaments. Using biochemical and immunological methods, these nuclear inclusion bodies were shown to be comprised of actin filaments. (b) Considerable alterations in the integrity of the nucleoli were observed after the heat-shock treatment. Specifically, there appeared to be a general relaxation in the condensation state of the nucleoli, changes in both the number and size of the granular ribonucleoprotein components, and finally a reorganization of the nucleolar fibrillar reticulum. These morphological changes in the integrity of the nucleoli are of significant interest since previous work as well as studies presented here show that two of the mammalian stress proteins, the major stress-induced 72-kD protein and the 110-kD protein, localize within the nucleoli of the cells after heat-shock treatment. We discuss these morphological changes with regards to the known biological and biochemical events that occur in cells after induction of the stress response.

https://rupress.org/jcb/article-pdf/101/4/1198/1052135/1198.pdf

Morphological study of the mammalian stress response: characterization of changes in cytoplasmic organelles, cytoskeleton, and nucleoli, and appearance of intranuclear actin filaments in rat fibroblasts after heat-shock treatment.

The relevant literature is reviewed in an attempt to clarify the mechanism of heat-dependent tumor cell destruction in vivo. Malignant cells in vivo appear to be selectively destroyed by hyperthermia in the range of 41-43 degrees C. Heat evidently affects nuclear function, expressed by an inhibited RNA, DNA and protein synthesis and characteristic arrest or delay of cells in certain locations of the cell cycle. However, as these effects appear to be reversible and are observed in normal cells as well as malignant cells, they probably do not explain the hyperthermic induced selective in vivo destruction of malignant cells. Heat-induced cytoplasmic damage appears to be of more importance. Increased lysosomal activation is observed, and is further intensified by a relatively increased anaerobic glycolysis which develops selectively in tumor cells. A hypothesis is proposed and discussed which explains the marked and selective in vivo tumor cell destruction as a consequence of the enhancing effect on the cytoplasmic damage of certain environmental factors (e.g. increased acidity, hypoxia and insufficient nutrition.

J.-P. Zalta

Papers

Effet de la température supra-optimale sur les ribonucléoprotéines et le RNA nucléolaire: I. Etude ultrastructurale☆

Effet de la température supra-optimale sur les ribonucléoprotéines et le RNA nucléolaire: II. Etude biochimique