scholarly journals Stage-specific patterns of collagen gene expression during development of Caenorhabditis elegans.

1985 ◽  
Vol 5 (2) ◽  
pp. 363-372 ◽  
Author(s):  
G N Cox ◽  
D Hirsh
Keyword(s):  
Caenorhabditis Elegans ◽  
Protein Composition ◽  
Large Family ◽  
Major Protein ◽  
Mrna Levels ◽  
Mrna Accumulation ◽  
C Elegans ◽  
Collagen Protein ◽  
Protein Components ◽  
Collagen Genes

Collagens are the major protein components of the Caenorhabditis elegans cuticle and are encoded by a large family of 40 to 150 closely related but nonidentical genes. We have determined temporal patterns of mRNA accumulation for a large number of collagen genes by screening recombinant phages and plasmids containing cloned collagen genes under high stringency conditions with 32P-labeled cDNA preparations specific for eggs or three postembryonic molts. We find that collagen mRNA levels are regulated both temporally and quantitatively during C. elegans development. Most genes studied exhibit one of four patterns of mRNA accumulation which correlate with changes in cuticle morphology and collagen protein composition during development. Our results suggest that, in general, there is a progressive activation of new collagen genes during normal development.

Stage-specific patterns of collagen gene expression during development of Caenorhabditis elegans

1985 ◽  
Vol 5 (2) ◽  
pp. 363-372
Author(s):  
G N Cox ◽  
D Hirsh
Keyword(s):  
Caenorhabditis Elegans ◽  
Protein Composition ◽  
Large Family ◽  
Major Protein ◽  
Mrna Levels ◽  
Mrna Accumulation ◽  
C Elegans ◽  
Collagen Protein ◽  
Protein Components ◽  
Collagen Genes

Collagens are the major protein components of the Caenorhabditis elegans cuticle and are encoded by a large family of 40 to 150 closely related but nonidentical genes. We have determined temporal patterns of mRNA accumulation for a large number of collagen genes by screening recombinant phages and plasmids containing cloned collagen genes under high stringency conditions with 32P-labeled cDNA preparations specific for eggs or three postembryonic molts. We find that collagen mRNA levels are regulated both temporally and quantitatively during C. elegans development. Most genes studied exhibit one of four patterns of mRNA accumulation which correlate with changes in cuticle morphology and collagen protein composition during development. Our results suggest that, in general, there is a progressive activation of new collagen genes during normal development.

scholarly journals Piceatannol, a Natural Stilbene, Extends the Lifespan of Caenorhabditis elegans Under Fasting Through Inhibition of Lipolysis

2020 ◽  
Vol 4 (Supplement_2) ◽  
pp. 38-38
Author(s):  
Jang Miran ◽  
Zhang Yuan ◽  
Bai Juan ◽  
Jun-Bae An ◽  
Park Yeonhwa ◽  
...  
Keyword(s):  
Lipid Metabolism ◽  
Caenorhabditis Elegans ◽  
Negative Energy ◽  
Hormone Sensitive Lipase ◽  
Pcr Analysis ◽  
Lifespan Extension ◽  
Mrna Levels ◽  
Free Glycerol ◽  
C Elegans ◽  
Glycerol Level

Abstract Objectives Lipolysis is the catabolic process that hydrolyzes triglyceride (TG) to free fatty acids (FFAs) and glycerol under negative energy balance such as fasting. In adipocytes, adipose TG lipase (ATGL), hormone-sensitive lipase (HSL), and monoglyceride lipase play key roles in a series of TG hydrolysis reactions in mammals. However, overly activated adipose lipolysis is believed to contribute to link between obesity and systemic inflammation and oxidative stress. We previously demonstrated that piceatannol (PIC), a natural resveratrol analogue, inhibits adipogenesis in cultured adipocytes and lipogenesis in Caenorhabditis elegans. Furthermore, we showed that PIC extends the lifespan of C. elegans via the insulin/IGF-1 signaling. However, the effects of PIC on lipid metabolism during fasting state is unknown. Methods We conducted Oil-Red-O assay, Enzyme assay (TG and Free glycerol contents), PCR analysis and lifespan assay. Results In this study, we demonstrated that PIC-treated C. elegans exhibited suppressed lipolysis under fasting as judged by increased lipid accumulation and TG levels with decreased free glycerol level. Consistent with these findings, PIC treatment resulted in decreased mRNA levels of genes involved lipolysis such as atgl-1, hosl-1 and aak-2 in fasted C. elegans. Also, PIC treatment augmented fasting-induced lifespan of C. elegans by an increased daf-16 gene expression. However, such effect was abolished when atgl-1, aak-2, and daf-16 mutants were treated with PIC. In addition, we also found that autophagy is required for PIC-induced lifespan in C. elegans during fasting since autophagy inhibitor treatments and autophagy gene deficient worms resulted in blunting the lifespan extension effect of PIC. Conclusions Collectively, our results indicate that PIC contributes to lifespan extension in C. elegans during fasting possibly through regulating lipolysis- and/or autophagy-dependent lipid metabolism. Funding Sources 1. The National Research Foundation of Korea (NRF) grant funded by the Korea government (MSIP) (2019R1A2C1086146) and (2019R1A6A3A03033878) 2. The Rural Development Administration of the Republic of Korea.

scholarly journals Venomics and Cellular Toxicity of Thai Pit Vipers (Trimeresurus macrops and T. hageni)

Toxins ◽  
2020 ◽  
Vol 12 (1) ◽  
pp. 54 ◽  
Author(s):  
Supeecha Kumkate ◽  
Lawan Chanhome ◽  
Tipparat Thiangtrongjit ◽  
Jureeporn Noiphrom ◽  
Panithi Laoungboa ◽  
...  
Keyword(s):  
Snake Venom ◽  
Protein Composition ◽  
Treatment Method ◽  
Cross Reactivity ◽  
Major Protein ◽  
Dependent Manner ◽  
Crude Venom ◽  
Monocytic Cells ◽  
Protein Components ◽  
Species Specific

The two venomous pit vipers, Trimeresurus macrops and T. hageni, are distributed throughout Thailand, although their abundance varies among different areas. No species-specific antivenom is available for their bite victims, and the only recorded treatment method is a horse antivenom raised against T. albolabris crude venom. To facilitate assessment of the cross-reactivity of heterologous antivenoms, protein profiles of T. macrops and T. hageni venoms were explored using mass-spectrometry-based proteomics. The results show that 185 and 216 proteins were identified from T. macrops and T. hageni venoms, respectively. Two major protein components in T. macrops and T. hageni venoms were snake venom serine protease and metalloproteinase. The toxicity of the venoms on human monocytes and skin fibroblasts was analyzed, and both showed a greater cytotoxic effect on fibroblasts than monocytic cells, with toxicity occurring in a dose-dependent rather than a time-dependent manner. Exploring the protein composition of snake venom leads to a better understanding of the envenoming of prey. Moreover, knowledge of pit viper venomics facilitates the selection of the optimum heterologous antivenoms for treating bite victims.

scholarly journals L-Ascorbate Biosynthesis Involves Carbon Skeleton Rearrangement in the Nematode Caenorhabditis elegans

Metabolites ◽  
2020 ◽  
Vol 10 (8) ◽  
pp. 334
Author(s):  
Yukinori Yabuta ◽  
Ryuta Nagata ◽  
Yuka Aoki ◽  
Ayumi Kariya ◽  
Kousuke Wada ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Caenorhabditis Elegans ◽  
Biosynthetic Pathway ◽  
Carbon Skeleton ◽  
Gas Chromatography Mass Spectrometry ◽  
Knockout Mutant ◽  
Nematode Caenorhabditis Elegans ◽  
C Elegans ◽  
Collagen Protein ◽  
Ascorbate Biosynthesis

Ascorbate (AsA) is required as a cofactor and is widely distributed in plants and animals. Recently, it has been suggested that the nematode Caenorhabditis elegans also synthesizes AsA. However, its biosynthetic pathway is still unknown. To further understand AsA biosynthesis in C. elegans, we analyzed the incorporation of the 13C atom into AsA using gas chromatography-mass spectrometry (GC-MS) in worms fed with D-Glc (1-13C)-labeled Escherichia coli. GC-MS analysis revealed that AsA biosynthesis in C. elegans, similarly to that in mammalian systems, involves carbon skeleton rearrangement. The addition of L-gulono-1,4-lactone, an AsA precursor in the mammalian pathway, significantly increased AsA level in C. elegans, whereas the addition of L-galactono-1,4-lactone, an AsA precursor in the plant and Euglena pathway, did not affect AsA level. The suppression of E03H4.3 (an ortholog of gluconolactonase) or the deficiency of F54D5.12 (an ortholog of L-gulono-1,4-lactone oxidase) significantly decreased AsA level in C. elegans. Although N2- and AsA-deficient F54D5.12 knockout mutant worm (tm6671) morphologies and the ratio of collagen to non-collagen protein did not show any significant differences, the mutant worms exhibited increased malondialdehyde levels and reduced lifespan compared with the N2 worms. In conclusion, our findings indicate that the AsA biosynthetic pathway is similar in C. elegans and mammals.

scholarly journals Number and organization of collagen genes in Caenorhabditis elegans.

1984 ◽  
Vol 4 (11) ◽  
pp. 2389-2395 ◽  
Author(s):  
G N Cox ◽  
J M Kramer ◽  
D Hirsh
Keyword(s):  
Caenorhabditis Elegans ◽  
Southern Blot ◽  
Blot Hybridization ◽  
Southern Blot Hybridization ◽  
Phage Library ◽  
Collagen Gene ◽  
Recombinant Phage ◽  
C Elegans ◽  
Dna Libraries ◽  
Collagen Genes

We analyzed the number and organization of collagen genes in the nematode Caenorhabditis elegans. Genomic Southern blot hybridization experiments and recombinant phage library screenings indicated that C. elegans has between 40 and 150 distinct collagen genes. A large number of recombinant phages containing collagen genes were isolated from C. elegans DNA libraries. Physical mapping studies indicated that most phage contained a single small collagen gene less than 3 kilobases in size. A few phage contained multiple collagen hybridizing regions and may contain a larger collagen gene or several tightly linked small collagen genes. No overlaps were observed between phages containing different collagen genes, implying that the genes are dispersed in the C. elegans genome. Consistent with the small size of most collagen genes, we found that the predominant class of collagen mRNA in C. elegans is 1.2 to 1.4 kilobases in length. Genomic Southern blot experiments under stringent hybridization conditions revealed considerable sequence diversity among collagen genes. Our data suggest that most collagen genes are unique or are present in only a few copies.

scholarly journals ANALYSIS OF CHD-7 DEFECTIVE DAUER NEMATODES IMPLICATES COLLAGEN MISREGULATION IN CHARGE SYNDROME FEATURES

2021 ◽  
Author(s):  
Diego Martín Jofré ◽  
Dane Kristian Hoffman ◽  
Ailen S. Cervino ◽  
McKenzie Grundy ◽  
Sijung Yun ◽  
...  
Keyword(s):  
Charge Syndrome ◽  
Mrna Levels ◽  
Craniofacial Malformations ◽  
Developmental Defects ◽  
The Public ◽  
Over Expression ◽  
C Elegans ◽  
Genetic Epistasis ◽  
Collagen Genes ◽  
Craniofacial Defects

ABSTRACTCHARGE syndrome is a complex developmental disorder caused by mutations in the chromodomain helicase DNA-binding protein7 (CHD7) and characterized by retarded growth and malformations in the heart and nervous system. However, despite the public health relevance of this disorder, relevant targets of CHD7 that relate to disease pathology are still poorly understood. Here we report thatchd-7, the nematode ortholog of CHD7, is required for dauer morphogenesis, lifespan determination, and stress response. Genetic epistasis placedchd-7in the TGF-β pathway. Consistent with our discoveries, we foundchd-7to be allelic toscd-3, a previously identified dauer suppressor from the TGF-β pathway. Interestingly, DAF-12 transcriptionally upregulatedchd-7, which is necessary to repressdaf-9for execution of the dauer program. Transcriptomic analysis comparingchd-7–defective and normal dauers showed enrichment of collagen genes, consistent with a conserved role for the TGF-β pathway in expression of the extracellular matrix. To validate a conserved function forchd-7in vertebrates, we usedXenopus laevisembryos, an established model to study craniofacial development. Morpholino mediated knockdown of Chd7 led to embryonic lethality, a reduction incol2a1mRNA levels and craniofacial defects in tadpoles. Both lethality and malformations were partially rescued in Chd7-depleted embryos by over-expression ofcol2a1. We suggest that pathogenic features of CHARGE syndrome caused by Chd7 mutations, such as craniofacial malformations, result from the reduction of collagen levels. These studies establishC. elegansas an amenable animal model to study the etiology of the developmental defects associated with pathogenic Chd7.

Molecular cloning and sequencing of ama-1, the gene encoding the largest subunit of Caenorhabditis elegans RNA polymerase II

1989 ◽  
Vol 9 (10) ◽  
pp. 4119-4130
Author(s):  
D M Bird ◽  
D L Riddle
Keyword(s):  
Caenorhabditis Elegans ◽  
Rna Polymerase ◽  
Rna Polymerase Ii ◽  
Rna Polymerase Iii ◽  
Gene Encoding ◽  
C Elegans ◽  
A Subunit ◽  
Rnap Ii ◽  
Collagen Genes ◽  
Splice Junctions

Two genomic sequences that share homology with Rp11215, the gene encoding the largest subunit of RNA polymerase II in Drosophila melanogaster, have been isolated from the nematode Caenorhabditis elegans. One of these sequences was physically mapped on chromosome IV within a region deleted by the deficiency mDf4, 25 kilobases (kb) from the left deficiency breakpoint. This position corresponds to ama-1 (resistance to alpha-amanitin), a gene shown previously to encode a subunit of RNA polymerase II. Northern (RNA) blotting and DNA sequencing revealed that ama-1 spans 10 kb, is punctuated by 11 introns, and encodes a 5.9-kb mRNA. A cDNA clone was isolated and partially sequenced to confirm the 3' end and several splice junctions. Analysis of the inferred 1,859-residue ama-1 product showed considerable identity with the largest subunit of RNAP II from other organisms, including the presence of a zinc finger motif near the amino terminus, and a carboxyl-terminal domain of 42 tandemly reiterated heptamers with the consensus Tyr Ser Pro Thr Ser Pro Ser. The latter domain was found to be encoded by four exons. In addition, the sequence oriented ama-1 transcription with respect to the genetic map. The second C. elegans sequence detected with the Drosophila probe, named rpc-1, was found to encode a 4.8-kb transcript and hybridized strongly to the gene encoding the largest subunit of RNA polymerase III from yeast, implicating rpc-1 as encoding the analogous peptide in the nematode. By contrast with ama-1, rpc-1 was not deleted by mDf4 or larger deficiencies examined, indicating that these genes are no closer than 150 kb. Genes flanking ama-1, including two collagen genes, also have been identified.

scholarly journals MLT-10 Defines a Family of DUF644 and Proline-rich Repeat Proteins Involved in the Molting Cycle of Caenorhabditis elegans

2010 ◽  
Vol 21 (10) ◽  
pp. 1648-1661 ◽  
Author(s):  
Vijaykumar S. Meli ◽  
Beatriz Osuna ◽  
Gary Ruvkun ◽  
Alison R. Frand
Keyword(s):  
Caenorhabditis Elegans ◽  
Fusion Protein ◽  
Molecular Mechanisms ◽  
Hormone Receptors ◽  
Large Family ◽  
Loss Of Function ◽  
Molting Cycle ◽  
C Elegans ◽  
Dynamic Assembly ◽  
Specific Proteins

The molting cycle of nematodes involves the periodic synthesis and removal of a collagen-rich exoskeleton, but the underlying molecular mechanisms are not well understood. Here, we describe the mlt-10 gene of Caenorhabditis elegans, which emerged from a genetic screen for molting-defective mutants sensitized by low cholesterol. MLT-10 defines a large family of nematode-specific proteins comprised of DUF644 and tandem P-X2-L-(S/T)-P repeats. Conserved nuclear hormone receptors promote expression of the mlt-10 gene in the hypodermis whenever the exoskeleton is remade. Further, a MLT-10::mCherry fusion protein is released from the hypodermis to the surrounding matrices and fluids during molting. The fusion protein is also detected in strands near the surface of animals. Both loss-of-function and gain-of-function mutations of mlt-10 impede the removal of old cuticles. However, the substitution mutation mlt-10(mg364), which disrupts the proline-rich repeats, causes the most severe phenotype. Mutations of mlt-10 are also associated with abnormalities in the exoskeleton and improper development of the epidermis. Thus, mlt-10 encodes a secreted protein involved in three distinct but interconnected aspects of the molting cycle. We propose that the molting cycle of C. elegans involves the dynamic assembly and disassembly of MLT-10 and possibly the paralogs of MLT-10.

Protein composition and protein synthesis in the surface membranes of Schistosoma mansoni

Parasitology ◽  
1972 ◽  
Vol 65 (1) ◽  
pp. 55-69 ◽  
Author(s):  
J. R. Kusel
Keyword(s):  
Protein Synthesis ◽  
Calcium Chloride ◽  
Unit Area ◽  
Surface Membrane ◽  
Biological Significance ◽  
Protein Composition ◽  
Major Protein ◽  
Freezing And Thawing ◽  
Protein Components ◽  
Protein Incorporation

Saponin treatment of cercariae and schistosomula alters the surface membrane so that it may be sheared from the organism and isolated as fragments by centrifugation. Saponin-calcium chloride treatment or freezing and thawing of adult worms removes the surface membrane, which can be washed from the bodies and collected by centrifugation. The small quantities of material available necessitated the development of a sensitive technique for detecting the proteins in the membranes. The surface membranes were radioiodinated after butan-1-ol extraction and electrophoresed in polyacrylamide gel. Adult surfaces prepared by the saponin-calcium chloride and by the freezing and thawing technique had identical protein components, detected as gel-cut profiles or in autoradiographs. The quantity of a rapidly migrating PAS-positive amido black negative component was greater in the surfaces prepared by saponincalcium chloride than in the frozen and thawed surfaces. This component contains lipid, some of which may be glycolipid. It was largely absent from the surface membranes of cercariae and schistosomula. Cercarial surface membranes contained a major protein component which was absent from the surface membranes of schistosomula. Otherwise the surface membranes of the cercariae were identical to those of the schistosomula in their protein components. The rate of incorporation of freshly synthesized protein per unit area of surface membrane of schistosomula was very low in the first 5 days in culture, after which there was a very rapid increase to a maximum rate on the 15th day. After this time, the rate of protein synthesis decreased to a low level at 26 days. In these studies the activity per unit area was measured and this would not be expected to vary greatly during growth. The biological significance of the observed variation in protein incorporation into the membrane is unclear.

scholarly journals Expression of β-1,4-galactosyltransferases during Aging in Caenorhabditis elegans

Gerontology ◽  
2020 ◽  
Vol 66 (6) ◽  
pp. 571-581
Author(s):  
Jennifer Kremer ◽  
Cornelia Brendel ◽  
Elisabeth Karin Maria Mack ◽  
Hildegard Isolde Dietlinde Mack
Keyword(s):  
Caenorhabditis Elegans ◽  
Model Organism ◽  
Mrna Levels ◽  
Germline Stem Cells ◽  
Wild Type ◽  
Activity Increase ◽  
C Elegans ◽  
Targeted Analysis ◽  
Age Dependent ◽  
Plasma Activity

<b><i>Background:</i></b> Altered plasma activity of β-1,4-galac­tosyl­transferases (B4GALTs) is a novel candidate biomarker of human aging. B4GALT1 is assumed to be largely responsible for this activity increase, but how it modulates the aging process is unclear at present. <b><i>Objectives:</i></b> To determine how expression of B4GALT1 and other B4GALT enzymes changes during aging of an experimentally tractable model organism, <i>Caenorhabditis elegans</i>. <b><i>Methods:</i></b> Targeted analysis of mRNA levels of all 3 <i>C. elegans</i> B4GALT family members was performed by qPCR in wild-type and in long-lived <i>daf-2</i> (insulin/IGF1-like receptor)-deficient or germline-deficient animals. <b><i>Results:</i></b> <i>bre-4</i> (<i>B4GALT1/2/3/4</i>) is the only B4GALT whose expression increases during aging in wild-type worms. In addition, <i>bre-4</i> levels also rise during aging in long-lived <i>daf-2</i>-deficient worms, but not in animals that are long-lived due to the lack of germline stem cells. On the other hand, expression of <i>sqv-3 (B4GALT7)</i> and of <i>W02B12.11 (B4GALT5/6)</i> appears decreased or constant, respectively, in all backgrounds during aging. <b><i>Conclusions:</i></b> The age-dependent <i>bre-4</i> mRNA increase in <i>C. elegans</i> parallels the age-dependent B4GALT activity increase in humans and is consistent with <i>C. elegans</i> being a suitable experimental organism to define potentially conserved roles of B4GALT1 during aging.

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