Temporal and spatial expression patterns of the small heat shock (hsp16) genes in transgenic Caenorhabditis elegans.

The expression of the hsp16 gene family in Caenorhabditis elegans has been examined by introducing hsp16-lacZ fusions into the nematode by transformation. Transcription of the hsp16-lacZ transgenes was totally heat-shock dependent and resulted in the rapid synthesis of detectable levels of beta-galactosidase. Although the two hsp16 gene pairs of C. elegans are highly similar within both their coding and noncoding sequences, quantitative and qualitative differences in the spatial pattern of expression between gene pairs were observed. The hsp16-48 promoter was shown to direct greater expression of beta-galactosidase in muscle and hypodermis, whereas the hsp16-41 promoter was more efficient in intestine and pharyngeal tissue. Transgenes that eliminated one promoter from a gene pair were expressed at reduced levels, particularly in postembryonic stages, suggesting that the heat shock elements in the intergenic region of an hsp16 gene pair may act cooperatively to achieve high levels of expression of both genes. Although the hsp16 gene pairs are never constitutively expressed, their heat inducibility is developmentally restricted; they are not heat inducible during gametogenesis or early embryogenesis. The hsp16 genes represent the first fully inducible system in C. elegans to be characterized in detail at the molecular level, and the promoters of these genes should find wide applicability in studies of tissue- and developmentally regulated genes in this experimental organism.

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‘Promoter trapping’ in Caenorhabditis elegans

Development ◽

10.1242/dev.113.2.399 ◽

1991 ◽

Vol 113 (2) ◽

pp. 399-408 ◽

Cited By ~ 4

Author(s):

I.A. Hope

Keyword(s):

Caenorhabditis Elegans ◽

Expression Patterns ◽

Cell Types ◽

Histochemical Staining ◽

Lacz Gene ◽

Dna Fragments ◽

C Elegans ◽

Active Expression ◽

Beta Galactosidase

A screen of gene expression patterns has been developed for the nematode Caenorhabditis elegans. Promoter-reporter gene fusions were constructed in vitro by ligating C. elegans genomic DNA fragments upstream of a lacZ gene. Patterns of beta-galactosidase expression were examined by histochemical staining of C. elegans lines transformed with the constructs. beta-galactosidase expression depended on translational fusion, so constructs were assayed in large pools to expedite detection of the low proportion that were active. Expression in a variety of cell types and temporal patterns was observed with different construct pools. The most striking expression patterns were obtained when the beta-galactosidase activity was localized to subcellular structures by the C. elegans portion of the fusion protein. The active constructs of three selected pools were identified subsequently by an efficient combinatorial procedure. The genomic locations of the DNA fragments from the active constructs were determined and appear to define previously uncharacterized genetic loci.

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PES-1 is expressed during early embryogenesis in Caenorhabditis elegans and has homology to the fork head family of transcription factors

Development ◽

10.1242/dev.120.3.505 ◽

1994 ◽

Vol 120 (3) ◽

pp. 505-514 ◽

Cited By ~ 4

Author(s):

I.A. Hope

Keyword(s):

Transcription Factors ◽

Caenorhabditis Elegans ◽

Cell Lineage ◽

Embryonic Cell ◽

Early Embryogenesis ◽

C Elegans ◽

Promoter Trapping ◽

Fork Head ◽

Initiation Of Transcription ◽

Beta Galactosidase

Promoter trapping has identified a gene, pes-1, which is expressed during C. elegans embryogenesis. The beta-galactosidase expression pattern, directed by the pes-1/lacZ fusion through which this gene was cloned, has been determined precisely in terms of the embryonic cell lineage and has three components. One component is in a subset of cells of the AB founder cell lineage during early embryogenesis, suggesting pes-1 may be regulated both by cell autonomous determinants and by intercellular signals. Analysis of cDNA suggests pes-1 has two sites for initiation of transcription and the two transcripts would encode related but distinct proteins. The predicted PES-1 proteins have homology to the fork head family of transcription factors and therefore may have important regulatory roles in early embryogenesis.

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The noncanonical small heat shock protein HSP-17 from Caenorhabditis elegans is a selective protein aggregase

Journal of Biological Chemistry ◽

10.1074/jbc.ra119.011185 ◽

2020 ◽

Vol 295 (10) ◽

pp. 3064-3079 ◽

Cited By ~ 1

Author(s):

Manuel Iburg ◽

Dmytro Puchkov ◽

Irving U. Rosas-Brugada ◽

Linda Bergemann ◽

Ulrike Rieprecht ◽

...

Keyword(s):

Caenorhabditis Elegans ◽

Heat Shock ◽

Small Heat Shock Protein ◽

Independent Manner ◽

C Elegans ◽

Higher Eukaryotes ◽

Prolonged Heat ◽

Shock Proteins ◽

Excretory Organs

Small heat shock proteins (sHsps) are conserved, ubiquitous members of the proteostasis network. Canonically, they act as “holdases” and buffer unfolded or misfolded proteins against aggregation in an ATP-independent manner. Whereas bacteria and yeast each have only two sHsps in their genomes, this number is higher in metazoan genomes, suggesting a spatiotemporal and functional specialization in higher eukaryotes. Here, using recombinantly expressed and purified proteins, static light-scattering analysis, and disaggregation assays, we report that the noncanonical sHsp HSP-17 of Caenorhabditis elegans facilitates aggregation of model substrates, such as malate dehydrogenase (MDH), and inhibits disaggregation of luciferase in vitro. Experiments with fluorescently tagged HSP-17 under the control of its endogenous promoter revealed that HSP-17 is expressed in the digestive and excretory organs, where its overexpression promotes the aggregation of polyQ proteins and of the endogenous kinase KIN-19. Systemic depletion of hsp-17 shortens C. elegans lifespan and severely reduces fecundity and survival upon prolonged heat stress. HSP-17 is an abundant protein exhibiting opposing chaperone activities on different substrates, indicating that it is a selective protein aggregase with physiological roles in development, digestion, and osmoregulation.

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A Hot Water Extract of Sideritis scardica Prolongs Life Span and Enhances Heat Shock Resistance in Caenorhabditis elegans

Natural Product Communications ◽

10.1177/1934578x20917283 ◽

2020 ◽

Vol 15 (4) ◽

pp. 1934578X2091728

Author(s):

Yoshihiko Nishioka ◽

Seiya Nishikawa ◽

Toshiyuki Shibata

Keyword(s):

Heat Stress ◽

Caenorhabditis Elegans ◽

Stress Response ◽

Heat Shock ◽

Life Span ◽

Water Extract ◽

Hot Water ◽

Control Group ◽

C Elegans ◽

Hot Water Extract

Sideritis scardica is a Lamiaceae plant that is endemic to the alpine zone of the Balkan Peninsula. The tea of S. scardica has been handed down as a “tea of longevity” in the Rhodope region of Bulgaria for an unknown amount of time. In this study, we prepared a hot water extract of S. scardica (SHWE) and examined its effects on both life span and stress response in living tissue using Caenorhabditis elegans and its transgenic mutants. The life span of wild-type N2 worms was prolonged by approximately 15% at the SHWE concentration of 5 µg/mL and approximately 22% at the SHWE concentration of 50 µg/mL, as compared with the control group. The effect of SHWE on the expression of heat shock protein 16.2 (HSP-16.2) under heat stress was investigated using TJ375 worms, a transgenic mutant of C. elegans. In the TJ375 worms pretreated with SHWE, the fluorescence intensity of green fluorescent protein fluorescence, which indicates the expression of HSP-16.2, was significantly increased. In the assay using TJ356 worms, the worms pretreated with SHWE did not show the translocation of DAF-16, a forkhead transcription factor class O homolog, from the cytoplasm to nucleus under heat stress. Additionally, under heat stress, the pretreatment of SHWE improved the survival rate of GR1307 worms, a knockout mutant of daf-16. These results indicate that SHWE enhances HSP-16.2 expression through a stress-response pathway (eg, HSF-1 pathway) other than the DAF-16 pathway, resulting in a prolonged life span of C. elegans under heat stress.

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Electrophysiological Measures of Aging Pharynx Function in C. elegans Reveal Enhanced Organ Functionality in Older, Long-lived Mutants

The Journals of Gerontology Series A ◽

10.1093/gerona/glx230 ◽

2017 ◽

Vol 74 (8) ◽

pp. 1173-1179 ◽

Cited By ~ 7

Author(s):

Joshua Coulter Russell ◽

Nikolay Burnaevskiy ◽

Bridget Ma ◽

Miguel Arenas Mailig ◽

Franklin Faust ◽

...

Keyword(s):

Caenorhabditis Elegans ◽

Heat Shock ◽

Life Span ◽

Insulin Signaling ◽

Model System ◽

Wild Type ◽

Organ Function ◽

C Elegans ◽

Age Related ◽

The Relationship

Abstract The function of the pharynx, an organ in the model system Caenorhabditis elegans, has been correlated with life span and motility (another measure of health) since 1980. In this study, in order to further understand the relationship between organ function and life span, we measured the age-related decline of the pharynx using an electrophysiological approach. We measured and analyzed electropharyngeograms (EPG) of wild type animals, short-lived hsf-1 mutants, and long-lived animals with genetically decreased insulin signaling or increased heat shock pathway signaling; we recorded a total of 2,478 EPGs from 1,374 individuals. As expected, the long-lived daf-2(e1370) and hsf-1OE(uthIs235) animals maintained pharynx function relatively closer to the youthful state during aging, whereas the hsf-1(sy441) and wild type animals’ pharynx function deviated significantly further from the youthful state at advanced age. Measures of the amount of variation in organ function can act as biomarkers of youthful physiology as well. Intriguingly, the long-lived animals had greater variation in the duration of pharynx contraction at older ages.

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Tissue- and paralogue-specific functions of acyl-CoA-binding proteins in lipid metabolism in Caenorhabditis elegans

Biochemical Journal ◽

10.1042/bj20102099 ◽

2011 ◽

Vol 437 (2) ◽

pp. 231-241 ◽

Cited By ~ 26

Author(s):

Ida C. Elle ◽

Karina T. Simonsen ◽

Louise C. B. Olsen ◽

Pernille K. Birck ◽

Sidse Ehmsen ◽

...

Keyword(s):

Caenorhabditis Elegans ◽

Unsaturated Fatty Acids ◽

Expression Patterns ◽

High Specificity ◽

Tissue Expression ◽

Yeast Cells ◽

Loss Of Function ◽

C Elegans ◽

Eukaryotic Species ◽

Quadruple Mutant

ACBP (acyl-CoA-binding protein) is a small primarily cytosolic protein that binds acyl-CoA esters with high specificity and affinity. ACBP has been identified in all eukaryotic species, indicating that it performs a basal cellular function. However, differential tissue expression and the existence of several ACBP paralogues in many eukaryotic species indicate that these proteins serve distinct functions. The nematode Caenorhabditis elegans expresses seven ACBPs: four basal forms and three ACBP domain proteins. We find that each of these paralogues is capable of complementing the growth of ACBP-deficient yeast cells, and that they exhibit distinct temporal and tissue expression patterns in C. elegans. We have obtained loss-of-function mutants for six of these forms. All single mutants display relatively subtle phenotypes; however, we find that functional loss of ACBP-1 leads to reduced triacylglycerol (triglyceride) levels and aberrant lipid droplet morphology and number in the intestine. We also show that worms lacking ACBP-2 show a severe decrease in the β-oxidation of unsaturated fatty acids. A quadruple mutant, lacking all basal ACBPs, is slightly developmentally delayed, displays abnormal intestinal lipid storage, and increased β-oxidation. Collectively, the present results suggest that each of the ACBP paralogues serves a distinct function in C. elegans.

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The Caenorhabditis elegans homologue of the extracellular calcium binding protein SPARC/osteonectin affects nematode body morphology and mobility.

Molecular Biology of the Cell ◽

10.1091/mbc.4.9.941 ◽

1993 ◽

Vol 4 (9) ◽

pp. 941-952 ◽

Cited By ~ 77

Author(s):

J E Schwarzbauer ◽

C S Spencer

Keyword(s):

Extracellular Matrix ◽

Caenorhabditis Elegans ◽

Calcium Binding ◽

Fusion Gene ◽

Muscle Cells ◽

Gene Organization ◽

The Body ◽

Matrix Assembly ◽

Developmentally Regulated ◽

C Elegans

The extracellular matrix-associated protein, SPARC (osteonectin [Secreted Protein Acidic and Rich in Cysteine]), modulates cell adhesion and induces a change in cell morphology. SPARC expression in mammals is developmentally regulated and is highest at sites of extracellular matrix assembly and remodeling such as parietal endoderm and bone. We have isolated cDNA and genomic DNA clones encoding the Caenorhabditis elegans homologue of SPARC. The gene organization is highly conserved, and the proteins encoded by mouse, human, and nematode genes are about 38% identical. SPARC consists of four domains (I-IV) based on predicted secondary structure. Using bacterial fusion proteins containing nematode domain I or the domain IV EF-hand motif, we show that, like the mammalian proteins, both domains bind calcium. In transgenic nematodes expressing a SPARC-lacZ fusion gene, beta-galactosidase staining accumulated in a striated pattern in the more heavily stained muscle cells along the body. Comparison of the pattern of transgene expression to unc-54-lacZ animals demonstrated that SPARC is expressed by body wall and sex muscle cells. Appropriate levels of SPARC are essential for normal C. elegans development and muscle function. Transgenic nematodes overexpressing the wild-type SPARC gene were abnormal. Embryos were deformed, and adult hermaphrodites had vulval protrusions and an uncoordinated (Unc) phenotype with reduced mobility and paralysis.

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The Nc1/Endostatin Domain of Caenorhabditis elegans Type Xviii Collagen Affects Cell Migration and Axon Guidance

The Journal of Cell Biology ◽

10.1083/jcb.152.6.1219 ◽

2001 ◽

Vol 152 (6) ◽

pp. 1219-1232 ◽

Cited By ~ 111

Author(s):

Brian D. Ackley ◽

Jennifer R. Crew ◽

Harri Elamaa ◽

Tania Pihlajaniemi ◽

Calvin J. Kuo ◽

...

Keyword(s):

Cell Migration ◽

Caenorhabditis Elegans ◽

Axon Guidance ◽

Ectopic Expression ◽

Basement Membranes ◽

Protein Isoforms ◽

Developmentally Regulated ◽

C Elegans ◽

Collagen Xviii ◽

Nc1 Domain

Type XVIII collagen is a homotrimeric basement membrane molecule of unknown function, whose COOH-terminal NC1 domain contains endostatin (ES), a potent antiangiogenic agent. The Caenorhabditis elegans collagen XVIII homologue, cle-1, encodes three developmentally regulated protein isoforms expressed predominantly in neurons. The CLE-1 protein is found in low amounts in all basement membranes but accumulates at high levels in the nervous system. Deletion of the cle-1 NC1 domain results in viable fertile animals that display multiple cell migration and axon guidance defects. Particular defects can be rescued by ectopic expression of the NC1 domain, which is shown to be capable of forming trimers. In contrast, expression of monomeric ES does not rescue but dominantly causes cell and axon migration defects that phenocopy the NC1 deletion, suggesting that ES inhibits the promigratory activity of the NC1 domain. These results indicate that the cle-1 NC1/ES domain regulates cell and axon migrations in C. elegans.

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Efficient transcription of a Caenorhabditis elegans heat shock gene pair in mouse fibroblasts is dependent on multiple promoter elements which can function bidirectionally.

Molecular and Cellular Biology ◽

10.1128/mcb.6.9.3134 ◽

1986 ◽

Vol 6 (9) ◽

pp. 3134-3143 ◽

Cited By ~ 26

Author(s):

R J Kay ◽

R J Boissy ◽

R H Russnak ◽

E P Candido

Keyword(s):

Caenorhabditis Elegans ◽

Heat Shock ◽

Gene Pair ◽

Mammalian Cells ◽

Heat Shock Gene ◽

Promoter Strength ◽

Wild Type ◽

Kinase Gene ◽

Mouse Fibroblasts ◽

Bidirectional Transcription

A divergently transcribed pair of Caenorhabditis elegans hsp16 genes was introduced into mouse fibroblasts by stable transfection with vectors containing bovine papillomavirus plasmid maintenance sequences and a selectable gene. The hsp16 genes were transcriptionally inactive in the mouse cells under normal growth conditions and were strongly induced by heat shock or arsenite. In a cell line with 12 copies of the gene pair, there were estimated to be more than 10,000 hsp16 transcripts in each cell after 2 h of heat shock treatment. The hsp16 transcript levels were more than 100 times higher than those of a gene with a herpes simplex virus thymidine kinase gene promoter carried on the same vector. A single heat shock promoter element (HSE) could activate bidirectional transcription of the two hsp16 genes when placed between the two TATA elements, but the transcriptional efficiency was reduced 10-fold relative to that of the wild-type gene pair. Four overlapping HSEs positioned between the two TATA elements resulted in inducible bidirectional transcription at greater than wild-type levels. The number of HSEs can therefore be a major determinant of the promoter strength of heat-inducible genes in mammalian cells. Partial disruption of an alternating purine-pyrimidine sequence between the two hsp16 genes had no significant effect on their transcriptional activity.

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The two gene pairs encoding H2A and H2B play different roles in the Saccharomyces cerevisiae life cycle

Molecular and Cellular Biology ◽

10.1128/mcb.7.10.3473-3481.1987 ◽

1987 ◽

Vol 7 (10) ◽

pp. 3473-3481

Author(s):

D Norris ◽

M A Osley

Keyword(s):

Saccharomyces Cerevisiae ◽

Life Cycle ◽

Heat Shock ◽

Heat Shock Response ◽

Spore Germination ◽

Gene Pair ◽

The Other ◽

Gene Pairs ◽

Gene Sets ◽

Shock Response

We have isolated Saccharomyces cerevisiae mutants bearing deletions of one or the other of the two divergently transcribed gene pairs encoding H2A and H2B. The deletions produced diverse effects on the yeast life cycle. Deletion of TRT1, one of the H2A-H2B gene pair sets, affected mitotic growth, sporulation, spore germination, the heat shock response, and exit from the stationary phase; deletion of TRT2, the other H2A-H2B gene pair set, had negligible effects on these same processes. Using a genetic complementation assay, we found that the differential effects of the deletions could be attributed to two features of the gene sets: first, the expression of the TRT1 gene pair, but not the TRT2 gene pair, could compensate for the absence of its partner; second, the protein subtypes encoded by the two gene pairs appear to have different functions in the heat shock response.

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