Mains PE (1994) Worm Breeder's Gazette "Interactions Between mei-1 and the unc-116 Kinesin"

Mains PE

[Worm Breeder's Gazette, 1994]

Interactions Between mei-l and the unc-116 Kinesin Paul E. Mains, Dept. of Medical Biochemistry, University of Calgary, Calgary, Alberta, Canada

Vladimir V Bakaev et al. (2003) Worm Breeder's Gazette "An attempt to slow aging in C. elegans. 31. No positive effect of glycerol"

Lyudmila M Bakaeva, Vladimir V Bakaev

[Worm Breeder's Gazette, 2003]

The purpose of this study was to investigate the effect of glycerol in water solutions on the nematode life span. In this experiment glycerol was used in following dilutions: 100 g/L, 10 g/L, 1 g/L, 100 mg/L, 10 mg/L, 1 mg/L and 0.1 mg/L. Three adult animals (3-5 days old) were kept in microtitre wells containing 0.5 ml of liquid medium (with E. coli and without glycerol) during 4 hours, then they were discarded and newborn larvae were transferred in next wells (without glycerol in medium) every day (one worm in one well) beginning from third day. Then, beginning from 3rd day, these worms were transferred every day in next wells containing medium with glycerol in any concentration. This investigation was carried out in temperature +21C and in the darkness. The obtained results are presented in the following table.

Yandell MD et al. (1994) Worm Breeder's Gazette "ceg-1 is Very Similar to decapentaplegic."

Wood WB, Yandell MD

[Worm Breeder's Gazette, 1994]

CEG-I IS VERY SIMILAR TO DECAPENTAPLEGIC Mark Yandell and Bill Wood Department of MCD Biology, University of Colorado, Boulder We have cloned a TGF-beta homolog which we have provisionally called ceg-l for C. elegans growth factor-l. ceg-l is a member of the dpp/bmp2-4 subclassl of TGF-beta proteins (see PILEUP diagram) and is very similar to the Drosophila gene decapentaplegic (dpp) and the mouse gene GDF-5, the product of the brachypodism locus, alleles of which cause defects in limb morphologyv.2 We previously reported (WBG 12(5): 82) that ceg-l maps to chromosome V just to the left of the cluster. We have now mapped ceg-l to the genetic map by PCR from single dead embryos homozygous for either the deficiency sDf30 or the deficiency sDf20 and find that it maps to the cluster of lethals near dpy-l l which lie under both deficiencies. Developmental Northern blots show that ceg-l encodes a moderately rare transcript that is present in late embryos and L1 larvae. Sequencing of genomic and cDNA clones reveal that the ceg-l gene has eight introns and encodes a 1.7kb message which is trans-spliced to SL1. We are now preparing to inject heat-shock and LacZ contructs. We are also collaborating with Ronald Plasterk to obtain a Tc1 insertion in ceg-l. Kingsley, D. M., Genes and Development 8: 133-146, 1994. 2 Storm, E. E., Huynh, T. V., Copland, N. G., Jenkins, N. A., Kingsley, D. M., and Lee, S. Nature 368: 639-643. 1994.

Vladimir V Bakaev et al. (2003) Worm Breeder's Gazette "An attempt to slow aging in C. elegans. 32. No positive effect of peroxide"

Vladimir V Bakaev, Lyudmila M Bakaeva

[Worm Breeder's Gazette, 2003]

The purpose of this study was to investigate the effect of hydrogen peroxide in water solutions on the nematode life span. In this experiment hydrogen peroxide was used in following dilutions: 100 mg/L, 10 mg/L, 1 mg/L, 0.1 mg/L, 0.01 mg/L and 0.001 mg/L. Three adult animals (3-5 days old) were kept in microtitre wells containing 0.5 ml of liquid medium (with E. coli and without hydrogen peroxide) during 4 hours, then they were discarded and newborn larvae were transferred in next wells (without hydrogen peroxide in medium) every day (one worm in one well) beginning from third day. Then, beginning from 3rd day, these worms were transferred every day in next wells containing medium with hydrogen peroxide in any concentration. This investigation was carried out in temperature +21C and in the darkness. The obtained results are presented in the following table. Conclusion: If hydrogen peroxide solution was applied to C. elegans, it was not able to increase their mean longevity significantly in comparison with control.

Kenyon CJ et al. (1994) Worm Breeder's Gazette "A polyray Suppressor"

Kenyon CJ, Maloof JN

[Worm Breeder's Gazette, 1994]

A polyray-l suppressor Julin Maloof and Cynthia Kenyon, Dept of Biochemistry, U.C.S.F, San Francisco, CA 94143 The polyray-l (pry-l) gene is required to keep homeotic selector (HOM-C) genes repressed where their expression has not been initiated (see accompanying abstract and WBG 12(5):39). pry-l mutant worms have numerous homeotic transformations and are very sick, so we were intrigued by a plate of pry-l mutant worms which appeared much healthier than normal. I outcrossed the strain and found F2s with the full pry-l mutant phenotype, suggesting that the "healthy" strain contained both pry-l and an extragenic suppressor. Does the suppressor have a phenotype on its own? Since pry-l causes homeotic transformations due to ectopic HOM-C expression, it seemed possible that the suppressor would cause transformations in the opposite direction, similar to those seen in HOM-C loss of function mutants. Indeed some F2s from the outcross had descendants of the QL neuroblast in the anterior of the worms, a phenotype also caused by loss-of function mutations in the HOM-C gene mab-5. We are calling this newly identified gene spy-l for suppressor of polyray-l. spy-l seems to suppress all of the pry-l mutant-phenotypes: pry-l; spy-l double mutant worms do not have extra rays, are no longer Muv, are not Unc, and have anteriorly placed QR descendants. spy-l could suppress pry-l either by preventing ectopic HOM-C expression, or by preventing ectopically expressed HOM-C genes from having any effect. mab-5-lacZ expression was examined and found to be normal in the p1y-l; spy-l double, so wild-type spy-l is required for the ectopic HOM-C expression seen in pry-l mutant animals. The fact that there is a QL migration defect in worms mutant for spy-l alone suggests that spy-l may normally be required to turn on mab-5 in QL. We compared expression of mab-5-lacZ in wild- type and spy-l mutant worms at 3.5-5 hours and at 8-10 hours after hatching. At 3.5-5 hours 100% (n=20) of the mutant and 95% (n=20) of the wild-type worms had expression in QL. At 8- 10 hours 0% (n=30) of the mutant worms has expression of mab- 5 in the descendants of QL, but 87% (n=30) of the wild-type worms showed expression. It appears that spy-l is not required to initiate mab-5 expression in QL, but that it is required to maintain expression once it is initiated. It is possible that spy-l is required to maintain the ON state of a number of HOM-C genes, in a manner analogous to the brahma and trithorax genes of Drosophila. spy-l is located on X, in a region well covered by cosmids. Hopefully a molecular analysis of spy-l along with the isolation and study of more spy-l alleles will enable us to determine how similar the C. elegans and Drosophila maintenance systems are, and how spy-l and other maintenance genes are used to maintain and specify pattern.

Ray C et al. (1995) Worm Breeder's Gazette "Evidence for Proteolytic Processing of a Cuticle Collagen in a Plant-Parasitic Nematode"

Hussey RS, Ray C

[Worm Breeder's Gazette, 1995]

Collagen is the major structural component of nematode cuticles. In Caenorhabditis elegans, the N-terminal regions of cuticle collagen proteins contain the highly conserved motif, R-X-X-R, which has been predicted to contain a potential subtilisin-like proteolytic processing site required for cleavage of the collagen proteins and normal cuticle collagen function (Kramer, 1994, FASEB 8:329-336). Cuticles of adult females of the plant-parasitic nematode Meloidogyne incognita contain a 76 kDa collagen which comprises over 50% of the B- mercaptoethanol-soluble cuticular proteins (Reddigari et al., 1986, J. Nematol. 18:294-302). We determined the N-terminal amino acid sequence of this major collagen and found it to be identical to the predicted amino acid sequence, starting at amino acid number 66, of the M. incognita Lemmi 5 cDNA clone (Van der Eycken et al., 1994, Gene 151:237-242) (Fig.1). A putative subtilisin-like protease recognition site was found immediately upstream of the region of amino acid homology between LEMMI 5 and the N-terminal sequence of the 76 kDa collagen (Fig.1). Our data support previous speculation about the existence of this novel method of collagen maturation and provide further evidence that this mechanism has been conserved during nematode evolution. In addition to protein processing, the expression of the Lemmi 5 gene was transcriptionally regulated: Lemmi 5-specific transcripts were present in adult females but not in eggs or second-stage juveniles. Also, Lemmi 5 analogs were present only in four Meloidogyne species, but not in C. elegans, Heterodera glycines, or tomato. .. PREDICTED LEMMI5 AMINO ACID SEQUENCE .. 1M A T L V V M P Q L Y S Q I N D L N L R V R D G V Q A .. F R V N T D S A W N D L M E L Q V A V T P Q S K P R S * N P F Q S L YR Q K RS L P D Y C I C Q P L E I N79 S L P D Y C I C Q P L E I N ............................................................ CONFIRMED N-TERMINAL AMINO ACID SEQUENCE .............................................................. Figure 1. The partial predicted amino acid sequence (residues 1-79) of the Lemmi 5 collagen gene is shown in roman letters. The putative subtilisin-like protease recognition sequence, R-Q-K-R, is boxed and the first amino acid after the proteolytic cleavage site is indicated by an *. The empirically determined N-terminal sequence of the 76 kDa from adult female M. incognita is italicized.

Grant BD et al. (1994) Worm Breeder's Gazette "sel-1, a Negative Regulator of lin-12 and glp-1 Activity, Encodes a Novel Extracellular Protein."

Greenwald IS, Grant BD

[Worm Breeder's Gazette, 1994]

sel-l, a negative regulator of lin-12 and glp-l activity, encodes a novel extracellular protein. Barth Grant and Iva Greenwald HHMI, Columbia University Dept. of Biochemistry, New York, NY 10032

Hodgkin JA et al. (1993) Worm Breeder's Gazette "Caenorhabditis Genetic Map and Nomenclature E-Mailing List"

Hodgkin JA, O'Callaghan M

[Worm Breeder's Gazette, 1993]

Most of the points covered in the accompanying 'Nomenclature Guidelines -- New Recommendations' contribution were circulated by e-mail before the June 1993 meeting, to principal investigators with e-mail addresses, for their comments and reactions. This procedure appeared to be efficient and helpful. We wish to expand our e-mailing list for nomenclature issues, and for recommended methods of data submission. For this reason, we sent a message in August 1993 inviting other potentially interested parties to join this list, using the e-mail addresses from the 1993 WBG Subscriber Directory. We repeat this message below, for the benefit of readers who have e-mail access but did not receive our invitation. If you are interested, send us an e-mail message asking to be added to the list. From cgc@mrc-lmb.cam.ac.uk Dear WBG subscriber At present, CGC e-mail messages about methods of data submission, nomenclatorial issues and so on, are sent to all investigators with assigned lab codes (strain and allele designations) and known e-mail addresses. We would like to extend this mailing list to other interested parties who can be reached by e-mail. If you wish to be included on this expanded e-mailing list, please reply to this message, and you will be added to the list. Investigators already on the list need take no action. Regardless of whether you join the list, you will still be able to take note of any significant new recommendations, because these will be duly announced in issues of the Worm Yours sincerely, Jonathan Hodgkin Mary O'Callaghan

Otsuka AJ et al. (1991) Worm Breeder's Gazette "Comparison of the unc-104 Protein 'Motor' Domain with Members of the Kinesin Family of Proteins"

Tang LZ, Whiteaker K, Boontrakulpoontatwee P, Otsuka AJ, Zhang Y

[Worm Breeder's Gazette, 1991]

The unc-104 gene is capable of encoding a kinesin-related protein ( Otsuka et al., Neuron, in press). Comparison of the presumed unc-104 'motor' domain to the ATPase and microtubule-binding region of members of the kinesin family demonstrates the greatest sequence identity in the putative ATP-binding domain [GYN-TIFAYGQTGSGKTYTM-G] and in conserved elements of unknown function, some of which may be involved in microtubule-binding [GIIPR, VK-S(Y/F)-E(I/L)YNE, DLL, R-VA-T--N-- SSRSH-VF-I, SGKL-LVDLAGSE, GAEG-R--E--NINKSL--LG-VI-AL, HIPYR(D/E)SKLT- LLQ-SLGG, N--ET-STL-(F/Y)A-RAK--(R/K)]. In the figure below, residues that are identical in 5 or more sequences are listed below the compilation and asterisks mark residues that are identical in all sequences. [See Figure 1]

Hishida R et al. (1994) Worm Breeder's Gazette "hch-1 Rescued"

Ishihara T, Kondo K, Katsura I, Hishida R

[Worm Breeder's Gazette, 1994]

hch-l Rescued Ryuichi Katsural I )DNA Research Center, National Institute of Genetics. Mishima, Shizuoka-ken 411, Japan, 2)Department of Biophysics, Faculty of Science, Kyoto University, Kyoto 606, Japan and 3)Department of Bioengineering, Faculty of Engineering, Soka University, Hachioji, Tokyo 192, Japan Mutations in the gene hch-l cause defects in both hatching and cell migration (E. Hedgecock et al., Development 100, 365-3~2, 1987). First, hch-l mutants exhibit delayed hatching, because the mutant embryos cannot digest proteins in their own eggshell. Second, the descendants of the QL neuroblast of hch-l mutants fail to migrate posteriorly, but migrate anteriorly, instead. How does hch-l gene function in these quite different systems? We have isolated a new allele of hch-l, utllO, from a mul-6 strain in the course of screening larval lethal mutations that cause gross morphological abnormalities (I. Katsura et al., WBG 12-2 plO8). Recently we cloned a unique Tcl containing fragment (HindIII 3.0kbp) from the genomic DNA of hch-l (utllO). A YAC filter and some cosmid clones were analyzed with a DNA fragment flanking the Tcl as a probe. This probe hybridized to YAC clones Y42D5, YlSF6, Y52Cll, Y43C6 and cosmid clones C42E7, F40ElO, M86, KllE3, TlSAlO, F23Cl. The result is consistent with genetic map position of hch-l gene (near unc-3 X). Since the Tcl insertion site is located near either end of the cosmid clones, we did not try to rescue hch-l worms with the cosmids. Instead, we cloned two DNA fragments containing the Tcl insertion site, a l5kbp PstI-Clal fragment and a 9kbp SacI fragment, from the N2 genomic DNA. These clones together with pRF4 as a dominant Rol marker were injected into hch-l (utl 10) mutants, and the extrachromosomally transformed worms were irradiated with gamma ray to obtain integrative transformants. The integrative transformants containing the Pstl-Clal fragment or the Sacl fragment show no delay of hatching. We are presently trying to rescue with shorter DNA fragments. Total RNA isolated from mixed staged N2 worms was analyzed by Northern blot using the Sacl fragment as a probe. This experiment revealed two transcripts (roughly 3kb and O.5kb in length) in this region. We plan to determine if these RNAs are encoded by hch-l gene and to isolate hch-l cDNA.