Manuel J Munoz et al. (2005) International Worm Meeting "Characterization of the long-lived mutants liv-7 and liv-8"

Ana M Brokate-Llanos, Manuel J Munoz, Isabel Calmarza-Font, Jose M Monje

[International Worm Meeting, 2005]

Using a positive selection for thermotolerant mutants (described in Munoz et al. Genetics 2003 163:171-80) we have isolated a total of 78 survivors of the thermal stress selection, among them, 24 are long lived. The characterization of two of those mutants will be described here. Mutation in the liv-7 gene produces a 40<sym05>/o increase of longevity, this effect is dependent of the fork head transcription factor daf-16. Beside the longevity phenotype, liv-7, at 27C, shows a weak delay in development, a phenotype that is independent of daf-16. Using the delay in development phenotype as surrogate genetic marker, we have located liv-7 in a small interval of the chromosome X, where no other long-lived mutant has been identified, suggesting that liv-7 encodes for a new gene involved on regulation of ageing. liv-8 shows a more pleiotropic phenotype. At 16C shows severe migration defects and at 25C it is lethal, when eggs are incubated at 25C majority of the embryos die, and those that do not die, stop development before reaching adulthood. However, if they are shifted up from 16C to 25C when adults, they are long lived. Mapping and further characterization of those mutants are on going.

Manuel J Munoz et al. (2002) European Worm Meeting "Positive selection of mutants with increased resistance and longevity."

Manuel A Fidalgo, Donald L Riddle, Manuel J Munoz, Jose M Monje

[European Worm Meeting, 2002]

We developed selective conditions for long-lived mutants by subjecting the first larval stage to thermal stress. At the moment, we have isolated a total of 64 thermotolerant mutants, 50 of them exhibited increased longevity. All the known classes of longevity mutants affected in the dauer signaling pathway were found (Unc, Dyf and Daf-c). 7 of them fail to complement daf-2, 5 age- 1, and 22 unc-31 all of them are known genes involved in the regulation of longevity. We also isolated 2 long-lived alleles of dyf-2 and one of unc-13, those genes were previously identified by other reasons and not related to longevity before. Thirteen additional mutations identified at least four new genes. We call this new mutant class Liv mutants (Long lIved and viable after thermal stress). One of these mutants is liv-6. This mutant resembles the age-1 reference allele, with Daf-c phenotype at 27C, long-lived and similar levels of thermotolerance. age-1 encodes a phosphatidylinositol 3 kinase (PI3K). liv-6 was mapped to the left arm of chromosome I, in the region where the only PI3K adapter subunit worm homologue is located. Sequencing of this gene in the mutant strain and rescue experiments demonstrated that liv-6 encodes for the PI3K adapter subunit.

McGhee JD (1987) Biochemistry "Purification and characterization of a carboxylesterase from the intestine of the nematode Caenorhabditis elegans."

McGhee JD

[Biochemistry, 1987]

The major intestinal esterase from the nematode Caenorhabditis elegans has been purified to essential homogeneity. Starting from whole worms, the overall purification is 9000-fold with a 10% recovery of activity. The esterase is a single polypeptide chain of Mr 60,000 and is stoichiometrically inhibited by organophosphates. Substrate preferences and inhibition patterns classify the enzyme as a carboxylesterase (EC 3.1.1.1), but the physiological function is unknown. The sequence of 13 amino acid residues at the esterase N- terminus has been determined. This partial sequence shows a surprisingly high degree of similarity to the N-terminal sequence of two carboxylesterases recently isolated from Drosophila mojavensis [Pen, J., van Beeumen, J., & Beintema, J. J. (1986) Biochem. J. 238, 691-699].

Lucas-Rodriguez, Patricia A. et al. (2021) International Worm Meeting "Looking for a possible treatment for type III galactosemia"

Lucas-Rodriguez, Patricia A., Brokate-Llanos, Ana Ma, Garzon , Andres, Munoz , Manuel J.

[International Worm Meeting, 2021]

Galactose metabolism is universally conserved in all living organisms, including Caenorhabditis elegans and humans. Galactose is metabolized by three conserved enzymes that constitute the Leloir pathway. GALE participates in the third step of the galactose metabolism pathway, catalyzing the interconversion of UDP-galactose (UDP-gal) and UDP-glucose (UDP-glu) and in some species, including C.elegans and humans, the interconversion of UDP-N-acetylgalactosamine (UDP-galNAc) and UDP-N-acetylglucosamine (UDP-gluNAc). All of these UDP-sugars are required for the glycosylation of proteins and lipids. Type III galactosemia is a rare disorder caused by mutations in GALE, homologue to the C. elegans gale-1 gen. The symptoms are due to the accumulation of intermediary galactose metabolites and the reduction of UDP-galNAc, because differently to the others, this sugar is only synthesized by GALE. Therefore, patients with type III galactosemia are recommended to follow a galactose-restrictive diet and take UDP-galNAc to avoid UDP-galNAc deficiency. This treatment is not able to improve the psychomotor retardation of these patients; it only slightly improves some of the symptoms. Our team has isolated a Caenorhabditis elegans mutant in the GALE homologue gene (gale-1(pv18)) which can be used as a model for type III galactosemia. Like humans, this mutant has an increase of UDP-gal and reduction of UDP-galNAc under regular diet. They are also sensitive to galactose rich diet and exhibit multiple developmental defects. We have found that the phenotype of the mutant and expression of gale-1 gene can be modified by diet. We also observed that treatment with some sugars improves the phenotype of the type III galactose model. The result observed, if conserved in humans, could be of interest to improve the quality of life of patients with galactosemia type III.

Murakami S et al. (1999) Curr Biol "Life extension and stress resistance in Caenorhabditis elegans modulated by the tkr-1 gene."

Johnson TE, Murakami S

[Curr Biol, 1999]

In this Brief Communication, which appeared in the 14 September 1998 issue of Current Biology, the UV dose was reported erroneously. The dose reported was 20 J/m2 but the actual dose used was 0.4 J/cm2. Also, the gene formally referred to as tkr-1 has since been renamed old-1 (overexpression longevity determination).

Ramos CG et al. (2014) J Bacteriol "Retraction for Ramos et al., MtvR is a global small noncoding regulatory RNA in Burkholderia cenocepacia."

Feliciano JR, da Costa PJ, Ramos CG, Grilo AM, Leitao JH

[J Bacteriol, 2014]

Volume 195, no. 16, p. 35143523, 2013. A number of problems related to images published in this paper have been brought to our attention. Figure 1D contains duplicated images in lanes S and LE, and Fig. 4D and 6B contain images previously published in articles in this journal and in Microbiology and Microbial Pathogenesis, i.e., the following: C. G. Ramos, S. A. Sousa, A. M. Grilo, J. R. Feliciano, and J. H. Leitao, J. Bacteriol. 193:15151526, 2011. doi:10.1128/JB.01374-11. S. A. Sousa, C. G. Ramos, L. M. Moreira, and J. H. Leitao, Microbiology 156:896908, 2010. doi:10.1099/mic.0.035139-0. C. G. Ramos, S. A. Sousa, A. M. Grilo, L. Eberl, and J. H. Leitao, Microb. Pathog. 48:168177, 2010. doi: 10.1016/j.micpath.2010.02.006. Therefore, we retract the paper. We deeply regret this situation and apologize for any inconvenience to the editors and readers of Journal of Bacteriology, Microbial Pathogenesis, and Microbiology.

Nillegoda NB et al. (2015) Nature "Crucial HSP70 co-chaperone complex unlocks metazoan protein disaggregation."

Berynskyy M, Morimoto RI, Bukau B, Stengel F, Kirstein J, Szlachcic A, Arnsburg K, Stank A, Scior A, Nillegoda NB, Gao X, Guilbride DL, Aebersold R, Wade RC, Mayer MP

[Nature, 2015]

Protein aggregates are the hallmark of stressed and ageing cells, and characterize several pathophysiological states. Healthy metazoan cells effectively eliminate intracellular protein aggregates, indicating that efficient disaggregation and/or degradation mechanisms exist. However, metazoans lack the key heat-shock protein disaggregase HSP100 of non-metazoan HSP70-dependent protein disaggregation systems, and the human HSP70 system alone, even with the crucial HSP110 nucleotide exchange factor, has poor disaggregation activity in vitro. This unresolved conundrum is central to protein quality control biology. Here we show that synergic cooperation between complexed J-protein co-chaperones of classes A and B unleashes highly efficient protein disaggregation activity in human and nematode HSP70 systems. Metazoan mixed-class J-protein complexes are transient, involve complementary charged regions conserved in the J-domains and carboxy-terminal domains of each J-protein class, and are flexible with respect to subunit composition. Complex formation allows J-proteins to initiate transient higher order chaperone structures involving HSP70 and interacting nucleotide exchange factors. A network of cooperative class A and B J-protein interactions therefore provides the metazoan HSP70 machinery with powerful, flexible, and finely regulatable disaggregase activity and a further level of regulation crucial for cellular protein quality control.

Miller DM et al. (1992) Worm Breeder's Gazette "unc-4 LacZ Expression in A-Type Motor Neurons"

Miller DM, Niemeyer CJ

[Worm Breeder's Gazette, 1992]

unc-4 LacZ expression in A-type motor neurons David M. Miller and Charles J. Niemeyer, Dept. of Cell Biology, Duke Univ. Medical Ctr, Durham, NC 27710

Sommer RJ et al. (1994) Worm Breeder's Gazette "Evolution of Vulva-Formation: Part II: Species With a Central Vulva"

Sommer RJ, Sternberg PW

[Worm Breeder's Gazette, 1994]

Evolution of vulva-formation: Part II: Species with a central vulva Ralf J. Sommer & Paul W. Sternberg, California Institute of Technology, Division of Biology 156-29, Pasadena, CA 91125

Jun-ichi Aikawa (2001) International C. elegans Meeting "Molecular characterization of heparan sulfate/heparin N-deacetylase/N-sulfotransferase in worms"

Jun-ichi Aikawa

[International C. elegans Meeting, 2001]

Heparan sulfate binds and activates a large variety of growth factors, enzymes and extracellular matrix proteins. These interactions largely depend on the specific arrangement of sulfated moieties and uronic acid epimers within the chains. These oligosaccharide sequences are generated in a step-wise manner, initiated by the formation of a linkage tetrasaccharide which is then extended by copolymerization of alternating a1,4GlcNAc and b1,4GlcA residues. As the chains polymerize, they undergo a series of sulfation and epimerization reactions. The first set of modifications involves the removal of acetyl units from subsets of GlcNAc residues, and the addition of sulfate groups to the resulting free amino groups. These reactions are catalyzed by a family of enzymes designated as GlcNAc N-deacetylase/N-sulfotransferases (NDST), since they simultaneously. Four members of the family have been identified in vertebrates, with single orthologs present in Drosophila and C. elegans. We have revealed tissue-specific expression pattern and unique enzymatic properties of these four isozymes1,2). In fly, loss of NDST (sulfateless) results in unsulfated chains and defective signaling by multiple growth factors and morphogens. I reconstituted cDNA for worm NDST from EST clones and 5' RACE products. Enzymatic activities will be discussed. 1) Aikawa, J. & Esko, J. D J. Biol. Chem. 274, 2690-2695 (1999) 2) Aikawa, J., Grobe, K., Tsujimoto, M. & Esko, J. D J. Biol. Chem. 276, 5876-5882 (2001)