Waring DA et al. (1990) Worm Breeder's Gazette "Does pal-1 have a homeobox?"

Kenyon CJ, Madhani H, Waring DA

[Worm Breeder's Gazette, 1990]

The gene pal-1 is required in males for the generation of rays by the V6 cells. As we have reported, pal-1 stimulates the production of rays by overcoming cell signals from the T cell that would otherwise inhibit ray production. (The Pal-1 V6 phenotype is suppressed by ablation of T.) The effect of pal-1 is to stimulate the function of the gene mab-5 and inhibit the function of lin-22 in V6 or its descendants (Cell, in press Jan. 12, 1990). We have rescued pal-1 animals by injection of a 14 kb fragment of DNA that appears to contain ceh-3 [one of the scores of homeoboxes found in C. elegans, ( Burglin et al. Nature 341:239)]. pal-1 maps very close to ced-4 on the left arm of linkage group III. Twelve recombinants between unc-79 and dyp-17 did not separate ced-4 and pal-1. unc-79 and dpy-17 are 0.25 map units apart. Thus with 95% confidence we know that ced-4 and pal-1 are less than or equal to 0.05 map units apart. This suggested that the genes might be within 50 kb of each other (using the conservative estimate of 1 Mb per map unit). ced-4 has been cloned and mapped to a contig by Junying Yuan. We therefore sent to England for a series of cosmids in the region (Thank you John and Alan) and began injecting to look for pal-1 rescue. We injected several of these cosmids (we used the gene unc-31 and the unc- 31-bearing cosmid C14G10 as a coinjection marker) and obtained heritable but unstable rescue of the Pal-1 phenotype with the cosmid W05E6. According to the cosmid map W05E6 completely spans the cosmid AC10 from which Burglin et al. cloned the homeobox ceh-3. It didn't take a Ph.D. to realize that this would be a good place to start looking for the pal-1 gene. Using the published ceh-3 protein sequence as a guide, we designed a degenerate oligonucleotide that should recognize the ceh-3 sequence, and used it to probe Southern blots of W05E6. We were thereby able to generate a restriction map of the cosmid and localize the hybridization to a 1.8 kb BamHI/SalI fragment. (Since we have not sequenced the hybridizing sequence we can not say unequivocally that this is ceh-3). We then subcloned a 14 kb (KpnI partial) fragment that contains the 1.8 kb Bam/Sal fragment (pWK14). We injected this 14 kb subclone into pal-1 animals and rescued the Pal-1 phenotype. As noted above pal-1 males are missing rays in their fans. Rescued animals had wildtype fans. In addition we saw one animal (out of 5) with an extra ray located in the body of the animal about halfway between the distal end of the gonad and the tail. Our numbers are still quite small so we cannot yet say how frequent the production of ectopic rays is. The generation of such ectopic rays might be due to negative control element(s) that are missing from the pWK14 plasmid or to overexpression of the gene caused by high copy number of the injected DNA, etc. In conclusion we have localized the pal-1 gene to a 14 kb fragment with the ability to rescue the Pal-1 phenotype. This fragment probably contains the homeobox ceh-3. Thus the question remains: Does pal-1 have a homeobox? We cannot be certain, but it looks like it might. If so this would suggest that the activity of pal-1 somehow overcomes the effect of cell signals by influencing transcription.

Thomas JH et al. (1994) Worm Breeder's Gazette "lin-36, a Class B Synthetic Multivulva Gene, Encodes a Novel Protein"

Thomas JH, Horvitz HR

[Worm Breeder's Gazette, 1994]

lin-36, a Class B Synthetic Multivulva Gene, Encodes a Novel Protein Jeffrey H. Thomas and H. Robert Horvitz, HHMI, Dept. Biology, MIT, Cambridge, MA 02139, USA

Imadzu H et al. (1994) Worm Breeder's Gazette "Two Tropomyosins Expressed in Body Wall and the Third Did In Pharynx of Caenorhabditis elegans"

Kagawa H, Sakube Y, Imadzu H

[Worm Breeder's Gazette, 1994]

TWO TROPOMYOSINS EXPRBSSBD IN BODY WALL AND THE THIRD DID IN PHARYNX OF CAENORHABDDITIS ELEGANS. H. Imadzu, Y. Sakube and H. gagawa. Department of Biology, Faculty of Science, Okayama University, Okayama, 700 Japan.

Sakube Y et al. (1994) Worm Breeder's Gazette "The Genome Structure and Expression of Promoter/lacZ Fusion Genes of C. elegans Ryanodine Receptor"

Imadzu H, Sakube Y, Kagawa H

[Worm Breeder's Gazette, 1994]

The Genome Structure and Expression of Promoter/lacZ Fusion Genes of the C. elegans Ryanodine Receptor Y. Sakube, H. Imadzu and H. Kagawa, Laboratory of Molecular Biology, Faculty of Science, Okayama University, Okayama, 700 Japan C51918@JPNKUDPC

Beitel GJ et al. (1994) Worm Breeder's Gazette "An ACEDB For the Macintosh User Group"

Beitel GJ, Horvitz HR

[Worm Breeder's Gazette, 1994]

An ACEDB for the Macintosh User Group Greg Beitel (BEITEL@MIT.EDU) and H. R. Horvitz, DepL Biology, MIT, Cambndge, MA 02139, USA

Ishii N et al. (1984) Worm Breeder's Gazette "sensitivity of the embryo of C elegans to X-rays."

Suzuki K, Ishii N

[Worm Breeder's Gazette, 1984]

Eggs isolated by the method using hyperchlorite were irradiated with X-rays after incubation for various times. The hatched worms were grown on agar plates with E. coli as food and survivors were scored for the animals reaching adulthood at 3 days after irradiation. The eggs at 2 h after egg collection were most sensitive and the D37 (dose corresponding to 37% survival) was 500 rads. The D37 of the eggs at 0, 2 and 3 h were 900, 750 and 700 rads, respectively and at 4 to 11 h were in the range from 5,000 to 8.000 rads. The eggs at 2 h was constituted of 100 to 350 cells; the number corresponding to that for the eggs of Rhabditidae tokai at 5 h, the most sensitive stage. The results showed that the eggs during the first half of embryogenesis (the stage of cell division) is about ten times more sensitive to X-irradiation than the eggs during the second half (the stage of morphogenesis). It was assumed that X-rays might influence not only cell division but also cell-cell-interactions.

Chamberlain SH et al. (1994) Worm Breeder's Gazette "A Molecular Approach to Identify skn-1-Responsive Genes in the Early C. elegans Embryo"

Chamberlain SH, Bowerman BA

[Worm Breeder's Gazette, 1994]

A molecular approach to identify skn-1-responsive genes in the early C. elegans embryo. Stephen H. Chamberlain and Bruce Bowerman, Institute of Molecular Biology, University of Oregon, Eugene, OR 97403

Huang Y-J et al. (1994) Worm Breeder's Gazette "From Ascaris to C. elegans: A Way to Study Gene Structure and Function"

Tobler H, Huang Y-J, Mueller F

[Worm Breeder's Gazette, 1994]

FROM ASCARIS TO C. ELEGANS: A WAY TO STUDY GENE STRUCTURE AND FUNCTION Huang Y-J., Tobler H. and Muller F., Institute of Zoology, University of Pribourg, Perolles, CH-1700 Fribourg, Switzerland

Marra MA et al. (1990) Worm Breeder's Gazette "Cloning and Sequencing the C. elegans Na+/H+ Antiporter Gene"

Clark DV, Baillie DL, Marra MA, Prasad SS

[Worm Breeder's Gazette, 1990]

The cosmid C11F2 has been shown to rescue two alleles of let-56, s46 and s173, using the technique of germ line transformation (1). Two adjacent PstI subclones of this cosmid detect transcripts on Northern blots. Neither of these two PstI subclones have been shown to rescue let-56. One PstI subclone (fragment 'q', see 2) detects a 2.4 kb message. This PstI subclone was used to screen a lambda gt10 cDNA library obtained from B. Meyer. This resulted in the isolation of a 1.3 kb cDNA, found to be contained entirely within fragment q. This 1. 3 kb cDNA (called 'C2'; 2) was sequenced and found to contain a single long open reading frame of approximately 1200 nucleotides. This cDNA was incomplete, representing the 3' portion of the expected message (3) . When the inferred amino acid sequence of this single long open reading frame was used to search the translated EMBL DNA databank ( release 20), significant homology was detected with the human growth factor activatable Na+/H+ antiporter protein sequence (3;4). We have commenced sequencing genomic C. elegans Na+/H+ antiporter DNA contained within a 6.2 kb HindIII fragment (called 'FH6'; 2) subcloned from fragment q. This subclone contains all of the C2 cDNA homologous region, plus an additional 800 nucleotides 5' of the C2 homologous sequence. Preliminary results indicate that the 5' end of the C. elegans Na+/H+ antiporter is not entirely contained within the additional 800 nucleotides of genomic DNA immediately upstream of the C2 homologous sequence residing within FH6. Currently, we are attempting to subclone the 5' end of the C. elegans Na+/H+ antiporter gene from fragment q. In addition, we will screen a lambda zap cDNA library obtained from B. Barstead in order to recover a full length Na+/H+ antiporter cDNA. Questions we intend to address include: (1) whether let-56 encodes the C. elegans Na+/H+ antiporter gene and (2) whether the C. elegans Na+/H+ antiporter protein is regulated in a manner similar to its human equivalent.

Eric G et al. (1995) Worm Breeder's Gazette "MARINER-LIKE ELEMENTS IN THE ENTOMOPARASITIC NEMATODE HETERORHABDITIS BACTERIOPHORA (NEMATODA RHABDITIDA)"

Pierre A, Monique A, Christian L, Eric G

[Worm Breeder's Gazette, 1995]

The mariner transposable element is a small member of the transposable element class II. This element is thought to transpose through a DNA intermediate and shows short inverted terminal repeats (ITR) bordering the entire element. Originally described in Drosophila mauritiana, it has been identified in many other insect species representing ten insect orders. Mariner-like elements (MLE) have also recently been identified in the planarian Dugesia tigrina and in the nematode Caenorhabditis elegans. Using a polymerase chain reaction (PCR) strategy with degenerated oligonucleotides deduced from conserved sequences of the mariner central region, we have been able to detect the presence of MLE in the entomoparasitic nematode Heterorhabditis bacteriophora. These MLE appear to be either deleted forms or full-sized elements of 1279 bp with imperfect ITR of 30 bp. In a full-sized element, the transposase part codes for 358 amino acids including three stop codons and two frame shifts. Therefore, none of the sequences has an open reading frame encoding for a putative transposase. The H. bacteriophora sequences present a 5' transposase longer than those usually described in the Tc/mariner-like family (addition of 14 amino acids). Most of the MLE conserved mariner amino acids (Robertson H.M., 1995, J.Ins.Physiol., 41(2):99) are present and 90% of those missing result from a single nucleotidic mutation. Interestingly, the H. bacteriophora transposase shares more similarities with insect MLE transposase than with C. elegans one. Among the conserved H. bacteriophora amino acids, only 14% are typical C. elegans amino acids (compared with C. elegans PM and C. elegans ZK 370 MLE) while 28% are typical insect ones (compared with Drosophila mauritiana Mos 1 and Hyalophora cecropia MLE). When analysed with Genbank data system, the most important similarity has been found with a Carpelimus sp. mariner (Genbank accession number U04455). The H. bacteriophora MLE is 73% similar, at the nucleotidic level, to this coleopteran MLE, while it is only 52% similar to a C. elegans mariner. On another hand, the H. bacteriophora MLE ITR are more similar to C. elegans MLE ITR (up to 53%) than to insect ones (about 38%). The distribution of H. bacteriophora MLE has been studied by Southern blot analysis. All the tested H. bacteriophora isolates contain MLE, while they seem to be absent from the Steinernema genus, a very closed genus of entomopathogenic nematodes too. These results, together with the fact that H. bacteriophora is an entomoparasitic rhabditid, strongly suggest a case of trans-phyla horizontal transfer. studies are still carried out to further caracterize this new mariner element and to understand the phylogenetic relationships between the mariner transposable elements isolated so far.