Phosphorylation of algal centrin is rapidly responsive to changes in the external milieu

1990 ◽  
Vol 96 (3) ◽  
pp. 395-402
Author(s):  
V.E. Martindale ◽  
J.L. Salisbury
Keyword(s):  
Protein Synthesis ◽  
Heat Shock ◽  
Green Alga ◽  
External Environment ◽  
Force Generation ◽  
Ethanol Treatment ◽  
Flagellar Roots ◽  
Ph Shock ◽  
Tetraselmis Striata ◽  
Root Contraction

Centrin, a calcium-sensitive contractile phosphoprotein, is the major component of the striated flagellar roots of the flagellate green alga, Tetraselmis striata. Flagellar roots contract in response to elevated calcium levels. Data presented here indicate that the level of centrin phosphorylation is rapidly responsive to changes in the cell's external environment. Centrin is dephosphorylated in response to elevated calcium or to heat shock. An increase in centrin phosphorylation accompanies pH shock or ethanol treatment. These changes are compared with flagellar excision, flagellar root contraction, and protein synthesis under the same treatments. We conclude that under certain conditions phosphorylation/dephosphorylation of centrin and extension/contraction of the flagellar root are uncoupled. Possible role(s) of centrin phosphorylation independent of force generation in the flagellar root are discussed.

scholarly journals Flagellar root contraction and nuclear movement during flagellar regeneration in Chlamydomonas reinhardtii.

1987 ◽  
Vol 105 (4) ◽  
pp. 1799-1805 ◽  
Author(s):  
J L Salisbury ◽  
M A Sanders ◽  
L Harpst
Keyword(s):  
Flagellar Apparatus ◽  
Central Position ◽  
Free Calcium ◽  
Cell Models ◽  
Nuclear Movement ◽  
Flagellar Roots ◽  
Flagellar Regeneration ◽  
Basal Apparatus ◽  
Ph Shock ◽  
Root Contraction

When Chlamydomonas cells are deflagellated by pH shock or mechanical shear the nucleus rapidly moves toward the flagellar basal apparatus at the anterior end of the cell. During flagellar regeneration the nucleus returns to a more central position within the cell. The nucleus is connected to the flagellar apparatus by a system of fibers, the flagellar roots (rhizoplasts), which undergo a dramatic contraction that coincides with anterior nuclear movement. A corresponding extension of the root system, back to its preshock configuration is observed as the nucleus retracts to a central position. Anterior displacement of the nucleus and flagellar root contraction require free calcium in the medium. Nuclear movement and flagellar root contraction and extension are not sensitive to inhibitors of protein synthesis (cycloheximide), or drugs that influence either microtubules (colchicine) or actin-based microfilaments (cytochalasin D). Detergent-extracted cell models contract and extend their flagellar roots and move their nuclei in response to alterations of free calcium levels in the medium. Cycles of nuclear movement in detergent-extracted models require ATP to potentiate the contractile mechanism for subsequent calcium-induced contraction. Flagellar root contraction and nuclear movement in Chlamydomonas may be causally related to signaling of induction of flagellar precursor genes or to the transport of flagellar precursors or their messages to sites of synthesis or assembly near the basal apparatus of the cell.

scholarly journals Characterization of the thermotolerant cell. I. Effects on protein synthesis activity and the regulation of heat-shock protein 70 expression.

1988 ◽  
Vol 106 (4) ◽  
pp. 1105-1116 ◽  
Author(s):  
L A Mizzen ◽  
W J Welch
Keyword(s):  
Protein Synthesis ◽  
Heat Shock ◽  
Stress Proteins ◽  
Stress Protein ◽  
Shock Treatment ◽  
Heat Shock Treatment ◽  
Normal Medium ◽  
Growth Environment ◽  
Mammalian Cell Lines ◽  
C 30

Exposure of mammalian cells to a nonlethal heat-shock treatment, followed by a recovery period at 37 degrees C, results in increased cell survival after a subsequent and otherwise lethal heat-shock treatment. Here we characterize this phenomenon, termed acquired thermotolerance, at the level of translation. In a number of different mammalian cell lines given a severe 45 degrees C/30-min shock and then returned to 37 degrees C, protein synthesis was completely inhibited for as long as 5 h. Upon resumption of translational activity, there was a marked induction of heat-shock (or stress) protein synthesis, which continued for several hours. In contrast, cells first made thermotolerant (by a pretreatment consisting of a 43 degrees C/1.5-h shock and further recovery at 37 degrees C) and then presented with the 45 degrees C/30-min shock exhibited considerably less translational inhibition and an overall reduction in the amount of subsequent stress protein synthesis. The acquisition and duration of such "translational tolerance" was correlated with the expression, accumulation, and relative half-lives of the major stress proteins of 72 and 73 kD. Other agents that induce the synthesis of the stress proteins, such as sodium arsenite, similarly resulted in the acquisition of translational tolerance. The probable role of the stress proteins in the acquisition of translational tolerance was further indicated by the inability of the amino acid analogue, L-azetidine 2-carboxylic acid, an inducer of nonfunctional stress proteins, to render cells translationally tolerant. If, however, analogue-treated cells were allowed to recover in normal medium, and hence produce functional stress proteins, full translational tolerance was observed. Finally, we present data indicating that the 72- and 73-kD stress proteins, in contrast to the other major stress proteins (of 110, 90, and 28 kD), are subject to strict regulation in the stressed cell. Quantitation of 72- and 73-kD synthesis after heat-shock treatment under a number of conditions revealed that "titration" of 72/73-kD synthesis in response to stress may represent a mechanism by which the cell monitors its local growth environment.

Coordinate changes in heat shock element-binding activity and HSP70 gene transcription rates in human cells

1988 ◽  
Vol 8 (11) ◽  
pp. 4736-4744
Author(s):  
D D Mosser ◽  
N G Theodorakis ◽  
R I Morimoto
Keyword(s):  
Protein Synthesis ◽  
Amino Acid ◽  
Heat Shock ◽  
Gene Transcription ◽  
Elevated Temperatures ◽  
Binding Activity ◽  
Heat Shock Gene ◽  
Hsp70 Gene ◽  
Heat Shock Element ◽  
Amino Acid Analogs

Activation of human heat shock gene transcription by heat shock, heavy metal ions, and amino acid analogs required the heat shock element (HSE) in the HSP70 promoter. Both heat shock- and metal ion-induced HSP70 gene transcription occurred independently of protein synthesis, whereas induction by amino acid analogs required protein synthesis. We identified a HSE-binding activity from control cells which was easily distinguished by a gel mobility shift assay from the stress-induced HSE-binding activity which appeared following heat shock or chemically induced stress. The kinetics of HSP70 gene transcription paralleled the rapid appearance of stress-induced HSE-binding activity. During recovery from heat shock, both the rate of HSP70 gene transcription and stress-induced HSE-binding activity levels declined and the control HSE-binding activity reappeared. The DNA contacts of the control and stress-induced HSE-binding activities deduced by methylation interference were similar but not identical. While stable complexes with HSE were formed with extracts from both control and stressed cells in vitro at 25 degrees C, only the stress-induced complex was detected when binding reactions were performed at elevated temperatures.

Heat-shock-induced grey crescent formation in axolotl eggs and oocytes: the role of gravity

Development ◽  
1987 ◽  
Vol 100 (4) ◽  
pp. 599-609
Author(s):  
J.-C. Beetschen ◽  
J. Gautier
Keyword(s):  
Protein Synthesis ◽  
Heat Shock ◽  
Synergistic Action ◽  
Nuclear Factor ◽  
Crescent Formation ◽  
Animal Pole ◽  
Grey Crescent ◽  
Cytoskeletal Structure

Axolotl eggs were heat shocked (36.8°C, 10min) inside their jelly layers. Heat shock (HS) was shown to induce the precocious appearance of a grey crescent (GC) in a number of eggs immediately after fertilization (Benford & Namenwirth, 1974). It was also demonstrated that this phenomenon occurs in fertilized or artificially activated eggs only when they are shocked within 11/2h after spawning. The GC forms still later in heated unfertilized, nonactivated eggs. The role of the jelly layers is considered to be mechanical: a proportion of eggs is maintained in a tilted position until the egg is able to orient animal pole upwards under the influence of gravity as a late consequence of activation. The jelly layers are not essential if the eggs are artificially tilted or rotated during HS. GC formation can also be induced in in vitro maturing oocytes, provided they are tilted during HS. Gravity thus plays an essential role in the cytoplasmic rearrangements leading to HS-induced GC formation. Our results indicate a synergistic action between heat and gravity in this process. The cytological appearance of the GC formed in those experiments is that of a ‘Born's crescent’ with a conspicuous ‘vitelline wall’ (Pasteels, 1964). When oocytes are enucleated before maturation, HS has no effect on GC formation. A nuclear factor is therefore essential, as has been demonstrated in early GC formation induced by inhibitors of protein synthesis. Finally, incorporation of amino acids into oocyte proteins appears to be rapidly inhibited by HS (from 5 min). However, we cannot conclude that GC formation is in fact triggered by inhibition of protein synthesis. It is also likely that HS disrupts cytoskeletal structure, hence facilitating cytoplasmic rearrangements. Nevertheless, these results are in agreement with the scheme we recently proposed for GC formation in the rotated axolotl oocyte (Gautier & Beetschen, 1985).

scholarly journals Ribosome Profiling Reveals HSP90 Inhibitor Effects on Stage-Specific Protein Synthesis in Leishmania donovani

mSystems ◽  
2018 ◽  
Vol 3 (6) ◽  
Author(s):  
Eugenia Bifeld ◽  
Stephan Lorenzen ◽  
Katharina Bartsch ◽  
Juan-José Vasquez ◽  
T. Nicolai Siegel ◽  
...  
Keyword(s):  
Gene Expression ◽  
Protein Synthesis ◽  
Heat Shock ◽  
Heat Shock Protein ◽  
Leishmania Donovani ◽  
Specific Protein ◽  
Ribosome Profiling ◽  
Severe Illness ◽  
Content Type ◽  
Hsp90 Activity

ABSTRACT The 90-kDa heat shock protein (HSP90) of eukaryotes is a highly abundant and essential chaperone required for the maturation of regulatory and signal proteins. In the protozoan parasite Leishmania donovani, causative agent of the fatal visceral leishmaniasis, HSP90 activity is essential for cell proliferation and survival. Even more importantly, its inhibition causes life cycle progression from the insect stage to the pathogenic, mammalian stage. To unravel the molecular impact of HSP90 activity on the parasites’ gene expression, we performed a ribosome profiling analysis of L. donovani, comparing genome-wide protein synthesis patterns in the presence and absence of the HSP90-specific inhibitor radicicol and an ectopically expressed radicicol-resistant HSP90 variant. We find that ribosome-protected RNA faithfully maps open reading frames and represents 97% of the annotated protein-coding genes of L. donovani. Protein synthesis was found to correlate poorly with RNA steady-state levels, indicating a regulated translation as primary mechanism for HSP90-dependent gene expression. The results confirm inhibitory effects of HSP90 on the synthesis of Leishmania proteins that are associated with the pathogenic, intracellular stage of the parasite. Those include heat shock proteins, redox enzymes, virulence-enhancing surface proteins, proteolytic pathways, and a complete set of histones. Conversely, HSP90 promotes fatty acid synthesis enzymes. Complementing radicicol treatment with the radicicol-resistant HSP90rr variant revealed important off-target radicicol effects that control a large number of the above-listed proteins. Leishmania lacks gene-specific transcription regulation and relies on regulated translation instead. Our ribosome footprinting analysis demonstrates a controlling function of HSP90 in stage-specific protein synthesis but also significant, HSP90-independent effects of the inhibitor radicicol. IMPORTANCE Leishmania parasites cause severe illness in humans and animals. They exist in two developmental stages, insect form and mammalian form, which differ in shape and gene expression. By mapping and quantifying RNA fragments protected by protein synthesis complexes, we determined the rates of protein synthesis for >90% of all Leishmania proteins in response to the inhibition of a key regulatory protein, the 90-kDa heat shock protein. We find that Leishmania depends on a regulation of protein synthesis for controlling its gene expression and that heat shock protein 90 inhibition can trigger the developmental program from insect form to mammalian form of the pathogen.

Thermosensitization, heat shock protein synthesis and development of thermotolerance in M-14 human tumor cells subjected to step-down heating

1992 ◽  
Vol 31 (4) ◽  
pp. 323-332 ◽  
Author(s):  
Andrea Delpino ◽  
Francesco Paolo Gentile ◽  
Francesca Di Modugno ◽  
Marcello Benassi ◽  
Anna Maria Mileo ◽  
...  
Keyword(s):  
Protein Synthesis ◽  
Heat Shock ◽  
Heat Shock Protein ◽  
Tumor Cells ◽  
Human Tumor ◽  
Human Tumor Cells ◽  
Step Down ◽  
M 14
Sign in / Sign up

Export Citation Format

Share Document