Alterations in the pattern of gene expression following heat shock in the nematode Caenorhabditis elegans

1983 ◽  
Vol 61 (6) ◽  
pp. 480-487 ◽  
Author(s):  
Terry P. Snutch ◽  
David L. Baillie
Keyword(s):  
Caenorhabditis Elegans ◽  
Heat Shock ◽  
Elevated Temperatures ◽  
In Vitro Study ◽  
In Vitro Translation ◽  
Nematode Caenorhabditis Elegans ◽  
Stage Of Development ◽  
Heat Shock Responses

Exposure of the nematode Caenorhabditis elegans to elevated temperatures induces the preferential synthesis of eight major polypeptides of approximate molecular weights 81 000, 70 000, 41 000, 38 000, 29 000, 19 000, 18 000, and 16 000. In pulse-labelled worms these peptides first appear at 29 °C and continue to be synthesized up to lethal temperatures. They are heat inducible at every stage of development. While temperature elevation induces the synthesis of the heat-shock polypeptides, the in vivo synthesis of most other proteins present before heat shock is suppressed. In contrast, in vitro translation of mRNA from heat-shocked worms shows no alteration from the pattern of normal 20 °C mRNAs except for the appearance of the heat-shock mRNAs. An in vitro study of RNA from control and heat-shocked dauer larvae shows that this developmental variant possesses little translatable mRNA but, upon heat shock, synthesizes a set of messages corresponding to the heat-shock polypeptides. The low background of this system will be especially useful in the analysis and purification of heat-shock mRNA for molecular cloning experiments. Extensive similarities between the Drosophila and C. elegans heat-shock responses are shown, including homology between the 70-kdalton heat-shock genes of the two organisms.

scholarly journals In Vivo and in Vitro Evidence that the Four Essential Intermediate Filament (IF) Proteins A1, A2, A3 and B1 of the Nematode Caenorhabditis elegans Form an Obligate Heteropolymeric IF System

2003 ◽  
Vol 333 (2) ◽  
pp. 307-319 ◽  
Author(s):  
Anton Karabinos ◽  
Ekkehard Schulze ◽  
Jürgen Schünemann ◽  
David A.D. Parry ◽  
Klaus Weber
Keyword(s):  
Caenorhabditis Elegans ◽  
Intermediate Filament ◽  
Nematode Caenorhabditis Elegans

scholarly journals Characterizing the structure-function relationship of a naturally-occurring RNA thermometer

2017 ◽  
Author(s):  
Sarai Meyer ◽  
Julius B. Lucks
Keyword(s):  
Heat Shock ◽  
Rna Structure ◽  
Structural Changes ◽  
Structural Motif ◽  
Ribosome Binding ◽  
Heat Shock Responses ◽  
Function Relationship ◽  
Relationship Of

AbstractA wide number of bacteria have been found to govern virulence and heat shock responses using temperature-sensing RNAs known as RNA thermometers. A prime example is theagsAthermometer known to regulate the production of the AgsA heat shock protein inSalmonella entericausing a “fourU” structural motif. Using the SHAPE-Seq RNA structure-probing methodin vivoandin vitro, we found that the regulator functions by a subtle shift in equilibrium RNA structure populations that lead to a partial melting of the helix containing the ribosome binding site. We also demonstrate that ribosome binding to theagsAmRNA causes changes to the thermometer structure that appear to facilitate thermometer helix unwinding. These results demonstrate how subtle RNA structural changes can govern gene expression and illuminate the function of an important bacterial regulatory motif.

scholarly journals Effects of supplementation with heat shock cognate 17 KDA protein (HSC70) on in vitro bovine embryo viability

2021 ◽  
Author(s):  
Aimé Jazmín Garza Arredondo ◽  
Diana Elisa Zamora Ávila ◽  
Uziel Castillo Velásquez ◽  
Gustavo Moreno Degollado ◽  
José Fernando De La Torre Sánchez ◽  
...  
Keyword(s):  
Heat Shock ◽  
Blastocyst Stage ◽  
Vital Role ◽  
Control Group ◽  
Bovine Embryo ◽  
Bovine Embryos ◽  
Embryo Viability ◽  
Stage Of Development

Abstract Endogenous heat shock cognate 71 kDa protein (HSC70) has a vital role in early embryonic development. This study assessed the effects of exogenous HSC70 on bovine embryo development and expression of genes associated with apoptosis. Expression analyses of HSPA1A, HSPA8, Bcl-2, and Bax genes were performed in bovine embryos in vivo on day 7 of development. Subsequently, expression of HSPA1A and HSPA8 were associated with apoptotic genes (Bcl-2 and Bax) in cultured bovine embryos in vitro that were supplemented with various concentrations (0 or control group, 50, and 100 ng) of HSC70. The results indicated that the control group (0 ng) in vitro embryos had higher expression of HSPA8, Bax, and Bcl-2 genes, compared with the vivo embryos (P < 0.01). In vitro-produced embryos supplemented with 50 ng or 100 ng HSC70 had higher expression of HSPA1A, HSC70, Bcl-2, and Bax genes, compared with the control group (P < 0.01). Embryos supplemented with 100 ng had greater expression of the HSPA8 gene compared with the control group and the group supplemented with 50 ng. However, embryos supplemented with 50 ng had better characteristics (i.e., stage of development and quality) than the control and 100-ng groups. In conclusion, supplementation of in vitro culture medium with HSC70 promoted development to the blastocyst stage and improved blastocyst quality.

scholarly journals Gene expression in Acetabularia. III. Comparison of stained cytosolic proteins and in vivo and in vitro translation products

1983 ◽  
Vol 60 (1) ◽  
pp. 1-12
Author(s):  
R.L. Shoeman ◽  
G. Neuhaus ◽  
H.G. Schweiger
Keyword(s):  
Messenger Rna ◽  
In Vitro Translation ◽  
Cytosolic Proteins ◽  
Molecular Weights ◽  
Translational Activity ◽  
Stage Of Development ◽  
Extended Analysis ◽  
Species Specific

A comparison of stained cytosolic proteins, in vivo 80 S ribosome translation products and in vitro translation products of poly(A)+ RNA from three species of Acetabularia was performed after characterization of their molecular weights and isoelectric points via two-dimensional electrophoresis. A total of 803 stained proteins, and 121 in vivo and 77 in vitro translation products, representing the most abundant proteins in each category, were analysed. In interspecies comparisons, approximately 10% of the stained proteins were common to all three species and more than 50% were found to be species-specific. Approximately 25% of the in vivo translation products were common to all three species and more than 30% were found to be species-specific. The majority of the in vivo and in vitro translation products were detected by one or both of the other methods employed. Even though the analysis was limited to the most abundant proteins detected by each of the three methods and to one stage of development, the results suggest that the translation of some proteins is not regulated, that the in vivo translation of others, whose mRNA is present and translated in vitro, is turned off while the translation in vivo of others is enhanced relative to the total. This feature makes them candidates for stage-specific proteins. The results provide a firm basis for the extended analysis of the biological activity of heterologous messenger RNA in Acetabularia cytoplasm and for a more complete cataloguing of the mRNA population and translational activity at different stages in the development of Acetabularia.

scholarly journals In virto and in vivo Phosphorylation of a Coat Protein of Potato Virus X

2021 ◽  
Vol 83 (5) ◽  
pp. 76-81
Author(s):  
L.O. Maksymenko ◽  
◽  
N.Y. Parkhomenko ◽  
Keyword(s):  
Coat Protein ◽  
Protein Phosphorylation ◽  
Potato Virus ◽  
Molecular Mechanisms ◽  
Potato Virus X ◽  
In Vitro Translation ◽  
Nitrocellulose Filter ◽  
Stage Of Development

At the present stage of development of plant virology the study of molecular mechanisms of regulation, translation and replication of viral RNA is of great interest. Potato virus X (PVX) RNA in viral particles is not available for in vitro translation, but acquires the ability to be translated as a result of shell protein phosphorylation. The aim of our study was to investigate the conditions of phosphorylation of the PVX coat protein in in vitro and in vivo systems, as well as the effect of EDTA and CaCl2 on the phosphorylation in vitro. Methods. The PVX coat protein was obtained by the guanidine chloride method. The kinase activity of PVX protein in vitro was determined in a standard reaction mixture containing Mn2+ ions, 0.8 mM EDTA, and 2 micro Ci 32P ATP (3000 Ci/mM). Phosphorylation of the protein in vivo was carried out by immersing Datura stramonium leaves with symptoms of PVX infection in water containing К3PO4 32P. After isolation of PVX from the leaves, the viral coat protein was fractionated by SDS-PAAG electrophoresis. Fractions of the protein were transferred from the gel by contact manner on a nitrocellulose filter. The PVX coat protein was detected by immunoblotting using immunoglobulins to PVX coat protein and rabbit antibodies labeled with peroxidase. The inclusion of labeled phosphorus in the PVX protein was detected by radioautography. Results. The PVX coat protein was phosphorylated in vitro in a standard incubation medium containing (gamma -32P) ATP. In contrast, the PVX coat protein cannot be phosphorylated in the same conditions in the presence of (alpha-32P) ATP. In vivo phosphorylated PVX coat protein was detected by exposing nitrocellulose filter with immunoblot on X-ray film. Additionally, it was found that the presence of 10 mm EDTA and 10 mm CaCl2 inhibited the process of the PVX coat protein phosphorylation in vitro. Conclusions. The coat protein of potato virus X is able to phosphorylate in vitro and in vivo systems. The terminal ATP phosphate plays a major role in the phosphorylation of the PVX coat protein. The presence of EDTA and Ca2+ influences on the process of protein phosphorylation in vitro. These agents are able to inhibit the process of phosphorylation of the PVX coat protein. Thus, the phenomenon of phosphorylation of the PVX coat protein apparently indicates about its participation in the regulation of the virus reproduction in the infected cell.

scholarly journals Talin requires beta-integrin, but not vinculin, for its assembly into focal adhesion-like structures in the nematode Caenorhabditis elegans.

1996 ◽  
Vol 7 (8) ◽  
pp. 1181-1193 ◽  
Author(s):  
G L Moulder ◽  
M M Huang ◽  
R H Waterston ◽  
R J Barstead
Keyword(s):  
Caenorhabditis Elegans ◽  
Focal Adhesion ◽  
Focal Adhesions ◽  
The Body ◽  
Nematode Caenorhabditis Elegans ◽  
C Elegans ◽  
Beta Integrin ◽  
Cell Shapes

In cultured cells, the 230-kDa protein talin is found at discrete plasma membrane foci known as focal adhesions, sites that anchor the intracellular actin cytoskeleton to the extracellular matrix. The regulated assembly of focal adhesions influences the direction of cell migrations or the reorientation of cell shapes. Biochemical studies of talin have shown that it binds to the proteins integrin, vinculin, and actin in vitro. To understand the function of talin in vivo and to correlate its in vitro and in vivo biochemical properties, various genetic approaches have been adopted. With the intention of using genetics in the study of talin, we identified a homologue to mouse talin in a genetic model system, the nematode Caenorhabditis elegans. C. elegans talin is 39% identical and 59% similar to mouse talin. In wild-type adult C. elegans, talin colocalizes with integrin, vinculin, and alpha-actinin in the focal adhesion-like structures found in the body-wall muscle. By examining the organization of talin in two different C. elegans mutant strains that do not make either beta-integrin or vinculin, we were able to determine that talin does not require vinculin for its initial organization at the membrane, but that it depends critically on the presence of integrin for its initial assembly at membrane foci.

Effects of TSH and calcitriol on bone metabolism: in vivo and in vitro study

2014 ◽  
Author(s):  
Ivo Dumic-Cule ◽  
Dunja Rogic ◽  
Damir Jezek ◽  
Lovorka Grgurevic ◽  
Slobodan Vukicevic
Keyword(s):  
Bone Metabolism ◽  
In Vitro Study ◽  
Vitro Study
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