Bernard V et al. (1995) Worm Breeder's Gazette "The Vps34 homolog of C. elegans"

Bernard V, Mueller F, Roggo L

[Worm Breeder's Gazette, 1995]

Phosphatidylinositol (3,4,5)P3 is considered to be a novel 2nd messenger and has been found to accumulate subsequently to cell surface receptor stimulation. A family of enzymes catalysing the phosphorylation of phosphoinositides at the D-3 position has been identified and cloned. Catalytic subunits of the p110[[alpha]] and [[beta]] -subtypes have been found in tight association with regulatory p85 subunits containing an SH3, two SH2 and a BCR domain. Activation of this heterodimeric PtdIns 3-kinase occurs during its translocation to autophosphorylated growth factor receptors, where the SH2 domains of p85 specifically recognise phosphorylated YXXM motives. The p110[[gamma]], on the other hand, is activated by [[beta]][[gamma]]-subunits of G-proteins and seems to transduce signals from serpentine receptors. While the p110[[alpha]]-[[gamma]] accept PtdIns, PtdIns 4-P and PtdIns(4,5)P2 as substrates, homologs of the yeast Vps34 gene product utilise solely PtdIns to produce PtdIns 3-P . Vps34p has been found to be indispensable for the sorting of various proteins to the yeast vacuole and is associated with the Vps15p serine/threonine kinase1.

Bernard V et al. (1994) Worm Breeder's Gazette "Cloning of the Ribosomal Protein S19 in Caenorhabditis elegans"

Mueller F, Etter A, Tobler H, Bernard V

[Worm Breeder's Gazette, 1994]

The nematode Ascaris lumbricoides undergoes chromatin diminution which causes a loss of about 25% of the total genomic DNA from all somatic cells. We have identified a ribosomal protein (rp) gene (coding for ALEP1 = Ascaris lumbricoides eliminated protein 1) which is located within the germ-line specific material. ALEP1 shows strong homologies with the eukaryotic ribosomal protein S19 family (rpS19 ).2D-gel electrophoresis on germ-line and somatic purified ribosome fractions shows that the ALEP1 protein (rpS19 )is present only in the germ-line ribosomes. In the rat, rpS19 has been localized at the tip of the small ribosomal subunit. What is the function of rpS19 ?Since Ascaris is not suitable for genetic analyses, we have isolated rpS19 from the nematode Caenorhabditis elegans. We used RT-PCR with degenerate oligonucleotides to amplify an 81 nucleotide long fragment. The Barstead cDNA library was screened with this fragment and positive clones were isolated and sequenced. The C. elegans rpS19 cDNA shows only 65% homology at the nucleic acid level. The transcript is trans-spliced with the splice leader SL1 .The deduced amino acid sequence predicts a protein of 150 amino acids with a molecular weight of 17 Kd. The C. elegans rpS19 protein shows perfect homology with other eukaryotic rpS19 proteins in the conserved regions. Overall, the RpS19 protein from C. elegans has 80% homology with Ascaris ALEPl. RpS19 is located on the C. elegans chromosome IV. Experiments are in progress to clone the C. elegans RpS19 gene. The identification of a rpS19 mutant in C. elegans might indicate its function.

Bernard V et al. (1993) International C. elegans Meeting "The primary structures of four ribosomal proteins from Ascaris lumbricoides."

Tobler H, Etter A, Bernard V, Nilsen TW, Mueller F

[International C. elegans Meeting, 1993]

Bernard V et al. (1995) International C. elegans Meeting "CAENORHADITIS ELEGANS HAS AN ABUNDANT PROTEIN C-LEA HOMOLOGOUS TO STRESS PROTEINS IN PLANTS"

Pythoud C, Mueller F, Bernard V, Tobler H

[International C. elegans Meeting, 1995]

We looked for abundant messengers in C. elegans and found a great number of cDNAs by screening a mix-stage cDNA library. The full length 2.3 Kb cDNA has been sequenced. The transcript is trans-spliced with the splice leader SL1. The deduced amino acid sequence of this C. elegans cDNA gives a protein of 732 amino acids with strong homologies to the plant LEA (late embryogenesis abundant) proteins. We thus called it C-LEA. It contains 20 repeats of an 11-mer amino acid repeating unit similar to those found in group 3 LEA proteins.

Smith BS et al. (1990) Cryobiology "Cryopreservation of the entomogenous nematode parasite Steinernema feltiae (= Neoaplectana carpocapsae)."

Hodgson-Smith A, Popiel I, James ER, Smith BS, Minter DM

[Cryobiology, 1990]

Cryopreservation studies were conducted with the J1 juvenile and third stage infective juvenile (IJ) larval stages of the entomogenous parasitic nematode Steinernema feltiae (=Neoaplectana carpocapsae). The main parameters evaluated were (i) tolerance of the organisms to the cryoprotectants methanol, ethanediol, glycerol, and dimethyl sulfoxide; (ii) the incubation temperature and exposure period in cryoprotectant; and (iii) slow cooling (circa 1C min-1) vs rapid cooling (circa 5,100C min-1). The J1 stage was sensitive to all cryoprotectants at >20% (v/v). This sensitivity increased at 22 and 37C. Exsheathed IJ stage organisms tolerated exposure to 50% (v/v) ethanediol or Me2SO and 60% (v/v) methanol or glycerol. A proportion (3.4%) of J1s preincubated in 20% (v/v) methanol for 10 min at 0C survived slow cooling to -35C followed by plunge into liquid nitrogen. Preincubation in 60% (v/v) Me2SO for 45 sec at 0C followed by rapid cooling yielded a higher proportion (12.3%0 of surviving J1s. The infective IJ stage only survived rapid cooling. Preincubation for 20 min at 0C in either 60% (v/v) methanol or 45% glycerol, followed by rapid cooling at 5,100C min-1, yielded 30-34% viable IJs following thawing. These larvae were capable of further growth and development in vitro. The results suggest that the organisms are vitrified during the rapid cooling step. For routine maintenance of strains of S. feltiae and related

Kane PM (2006) Microbiol Mol Biol Rev "The where, when, and how of organelle acidification by the yeast vacuolar H+-ATPase."

Kane PM

[Microbiol Mol Biol Rev, 2006]

All eukaryotic cells contain multiple acidic organelles, and V-ATPases are central players in organelle acidification. Not only is the structure of V-ATPases highly conserved among eukaryotes, but there are also many regulatory mechanisms that are similar between fungi and higher eukaryotes. These mechanisms allow cells both to regulate the pHs of different compartments and to respond to changing extracellular conditions. The Saccharomyces cerevisiae V-ATPase has emerged as an important model for V-ATPase structure and function in all eukaryotic cells. This review discusses current knowledge of the structure, function, and regulation of the V-ATPase in S. cerevisiae and also examines the relationship between biosynthesis and transport of V-ATPase and compartment-specific regulation of acidification.

Goddard JM et al. (1986) Proc Natl Acad Sci U S A "Isolation and characterization of Caenorhabditis elegans DNA sequences homologous to the v-abl oncogene."

Goddard JM, Capecchi MR, Weiland JJ

[Proc Natl Acad Sci U S A, 1986]

DNA sequences homologous to the v-abl oncogene were isolated from a Caenorhabditis elegans genomic library by their ability to hybridize with a v-src probe. The DNA sequence of 2465 nucleotides of one clone was determined. This region corresponds to the 5' protein kinase domain of v-abl plus approximately equal to 375 base pairs toward the 3' end. Four potential introns were identified. The homology between the deduced amino acid sequence of the C. elegans clone and that of the 1.2-kilobase-pair protein kinase region of v-abl is 62%. The tyrosine residue corresponding to the tyrosine that is phosphorylated in the v-src protein is conserved in the C. elegans sequence. When 95 amino acids around this tyrosine were compared with the corresponding sequences of Drosophila c-abl, v-abl, and v-src, the identities were 83%, 79%, and 56%, respectively. Hybridization of the cloned DNA with C. elegans poly(A)+ RNA revealed a major transcript of 4.4 kilobases.

Ernstrom GG et al. (2012) Genetics "V-ATPase V1 sector is required for corpse clearance and neurotransmission in Caenorhabditis elegans."

Weimer R, Watanabe S, Pawar DR, Greenstein D, Ernstrom GG, Jorgensen EM, Hobson RJ

[Genetics, 2012]

The vacuolar-type ATPase (V-ATPase) is a proton pump composed of two sectors, the cytoplasmic V(1) sector that catalyzes ATP hydrolysis and the transmembrane V(o) sector responsible for proton translocation. The transmembrane V(o) complex directs the complex to different membranes, but also has been proposed to have roles independent of the V(1) sector. However, the roles of the V(1) sector have not been well characterized. In the nematode Caenorhabditis elegans there are two V(1) B-subunit genes; one of them, vha-12, is on the X chromosome, whereas spe-5 is on an autosome. vha-12 is broadly expressed in adults, and homozygotes for a weak allele in vha-12 are viable but are uncoordinated due to decreased neurotransmission. Analysis of a null mutation demonstrates that vha-12 is not required for oogenesis or spermatogenesis in the adult germ line, but it is required maternally for early embryonic development. Zygotic expression begins during embryonic morphogenesis, and homozygous null mutants arrest at the twofold stage. These mutant embryos exhibit a defect in the clearance of apoptotic cell corpses in vha-12 null mutants. These observations indicate that the V(1) sector, in addition to the V(o) sector, is required in exocytic and endocytic pathways.

Jennifer M Ross et al. (2002) Mid-west Worm Meeting "A role for the Polycomb Group in the development of the C. elegans male nervous system"

Jennifer M Ross, David A Zarkower

[Mid-west Worm Meeting, 2002]

Development of the C. elegans male-specific nervous system depends on interplay between two regulatory pathways, one involving the Hox genes mab-5 and egl-5, and the other involving the conserved sexual regulator mab-3. Males harboring Hox or mab-3 mutations lack V rays, sensory structures required for mating. The Hox proteins activate the bHLH gene lin-32 to specify the V rays, while the doublesex homolog MAB-3 synergizes with the Hox/lin-32 pathway to promote V ray differentiation. V rays can be restored in mab-3(-) males by overexpression of lin-32, suggesting that LIN-32 acts as a primary determinant of V ray fate, while MAB-3 potentiates LIN-32 activity.

Baird, Scott E. et al. (2011) International Worm Meeting "Ray Pattern Variation in C. elegans: Mapping a Major-Effect QTL on LGV."

Baird, Scott E., Bailey, Daniel

[International Worm Meeting, 2011]

Cryptic variation of ray pattern has been observed in recombinant-inbred lines (RIL) derived from crosses of N2 and CB4856 strains of C. elegans (Guess et al., 2007). The variant pattern consists of the anterior displacement of ray 3 into a position immediately adjacent to ray 2. QTL analyses of these RIL identified a major-effect QTL on the left arm of chromosome V (QTL-V). One RIL, QX34, possessed a recombinant chromosome V that contained CB4856 alleles from -12.72 to -7.93 cM, a region that precisely corresponded to QTL-V. When this chromosome (QX34-V) was crossed into an otherwise N2 background (strain PB2034), the variant ray pattern was observed at a frequency of 0.50. Based on genotypes of other RIL with recombination breakpoints in this region, it appeared that QTL-V resulted from allelic variation at two or more loci. To confirm this result, we are mapping the gene(s) in this region responsible for the observed variation in ray pattern. Initial experiments demonstrated that the CB4856 alleles responsible for QTL-V were recessive to the corresponding N2 alleles. From deletion heterozygotes, it was determined that at least one gene responsible for QTL-V lies to the right of -9.00 cM. However, deletion of a region from -12.06 to -8.15 did not uncover the ray pattern variant. Therefore, either allelic variation at two or more genes is required for QTL-V or the gene responsible for QTL-V resides between -8.15 and -7.93 cM. Attempts at recombination mapping of QTL-V are ongoing but have been hindered by an apparent suppression of recombination in PB2034/N2 heterozygotes.