Small roundworms such as Caenorhabditis elegans release chemical signals called ascarosides in order to communicate with other worms of the same species. Using the ascarosides, the worm can tell its friends, for example, how crowded the neighborhood is and whether there is enough food. The ascarosides thus help the worms in the population decide whether the neighborhood is good – meaning they should hang around, eat, and make babies – or whether the neighborhood is bad. If so, the worms should develop into a larval stage specialized for dispersal that will allow them to find a better neighborhood.

Roundworms make the ascarosides by attaching a long chemical ‘side chain’ to an ascarylose sugar. Further chemical modifications allow the worms to produce different signals. In general, to signal a good neighborhood, worms attach a structure called an indole group to the ascarosides. To signal a bad neighborhood, worms make the side chain very short. But how does a worm control which ascarosides it makes?

Zhou, Wang et al. now show that C. elegans can change the meaning of its chemical message by modifying the ascarosides that it has already produced instead of making new ones from scratch. Specifically, as their neighborhood runs out of food, C. elegans can use an enzyme called ACS-7 to initiate the shortening of the side chains of indole-ascarosides. The worm can thus change a favorable ascaroside signal that causes the worms to group together into an unfavorable ascaroside signal that causes the worms to enter their dispersal stage.

Although Zhou, Wang et al. have focused on chemical communication in C. elegans, the findings could easily apply to the many other species of roundworm that produce ascarosides. Knowing how worms communicate will help us to understand how worms respond to their environment. This knowledge could potentially be used to interfere with the lifecycles and survival of parasitic worm species that harm health and crops.

The nematode C. elegans secretes ascarosides as pheromones to induce development of the dauer larval stage at high population densities, as well as to control various behaviors, including male attraction to hermaphrodites, hermaphrodite attraction to males, avoidance, foraging behavior, and adult aggregation (Cassada and Russell, 1975; Golden and Riddle, 1982; Butcher et al., 2007; Butcher et al., 2008; Srinivasan et al., 2008; Butcher et al., 2009a; Pungaliya et al., 2009; Izrayelit et al., 2012; Srinivasan et al., 2012; Artyukhin et al., 2013; Greene et al., 2016). The ascarosides are derivatives of the 3,6-dideoxysugar ascarylose that can be named according to their structural features, including the number of carbons (C#) in their fatty acid-derived side chains (Butcher, 2017). These side chains are attached to the ascarylose sugar at their penultimate (ω−1) or terminal (ω) carbon and are often unsaturated (Δ) at the α-β position. The ascarosides can be modified with various head groups at the 4’-position of the ascarylose sugar, such as the IC, the octopamine succinyl (OS), the 4-hydroxybenzoyl (HB), and the (E)-2-methyl-2-butenoyl (MB) groups. They can also be modified with various terminus groups at the end of their fatty-acid side chain. These head and terminus modifications can have important consequences for the biological activity of the ascarosides. For example, while unmodified, simple ascarosides are often negative signals that induce avoidance in C. elegans, ascarosides that are modified with an IC head group, such as IC-asc-ΔC9 (icas#3), are favorable signals that induce aggregation at picomolar concentrations (Srinivasan et al., 2012; von Reuss et al., 2012). However, the consequences of the IC group and other modifications can be quite nuanced. For example, unlike IC-ascarosides with medium-length (C9) side chains, which do not induce dauer, an IC-ascaroside with a short (C5) side chain, IC-asc-C5 (icas#9), is one of the five primary components of the dauer pheromone and induces the dauer larval stage when it accumulates at low nanomolar concentrations (Butcher et al., 2009a). Thus, IC-ascarosides have a dual nature to their activity as they are generally favorable signals that induce aggregation on food, but can also indicate unfavorable conditions and induce dauer when they have short side chains and accumulate to higher concentrations.

Peroxisomal β-oxidation is central to biosynthesis of the ascarosides. β-oxidation cycles shorten the side chains of long-chain ascaroside precursors by two carbons per cycle to produce short- and medium-chain ascaroside pheromones (Golden and Riddle, 1985; Wanders and Waterham, 2006; Butcher et al., 2009b; Joo et al., 2009; Joo et al., 2010; von Reuss et al., 2012; Zhang et al., 2015; Zhang et al., 2018). The first step in these β-oxidation cycles is catalyzed by an acyl-CoA oxidase, which installs a double bond at the α-β position of its substrate in a reaction that uses an FAD cofactor and produces H₂O₂. Specific acyl-CoA oxidases process ascaroside precursors with specific side-chain lengths and thus help to control which pheromones are produced (Joo et al., 2010; von Reuss et al., 2012; Zhang et al., 2015; Zhang et al., 2018). The C. elegans genome encodes seven acyl-CoA oxidases, five of which have been implicated in ascaroside production (Zhang et al., 2018). To reflect their similarity to mammalian ACOX-1, the acyl-CoA oxidases that were previously referred to as ACOX-1, –2, −3, –4, and −5 (von Reuss et al., 2012; Zhang et al., 2015; 2016) have been renamed ACOX-1.1, –1.2, −1.3, –1.4, and −1.5, respectively, and an additional acyl-CoA oxidase (F59F4.1) has been named ACOX-1.6. To reflect its similarity to mammalian ACOX-3, the acyl-CoA oxidase that was previously referred to as ACOX-6 (Zhang et al., 2016) has been renamed ACOX-3. The remaining three steps in each β-oxidation cycle are catalyzed by an enoyl-CoA hydratase (MAOC-1), an (R)-3-hydroxyacyl-CoA dehydrogenase (DHS-28), and a 3-ketoacyl-CoA thiolase (DAF-22) (Butcher et al., 2009b; Joo et al., 2009; Joo et al., 2010; von Reuss et al., 2012; Zhang et al., 2015). Our group has shown that (ω−1)-ascarosides and ω-ascarosides are biosynthesized by two parallel β-oxidation pathways, the former pathway involving ACOX-1.1, ACOX-1.3, ACOX-1.4, and ACOX-3 and the latter pathway involving ACOX-1.1 and ACOX-1.2 (Zhang et al., 2015; Zhang et al., 2018). In general, extensive β-oxidation of ascaroside precursors by these pathways to make ascarosides with very short side chains generates some of the most potent dauer pheromones, such as asc-ωC3 (ascr#5).

Very little is known about how various head groups, such as the IC, OS, HB, and MB groups, become attached to the ascarosides and how this attachment process is coordinated with the β-oxidation process that shortens the ascaroside side chains. Attachment of the IC head group to the 4’-position of the ascarylose is thought to occur late in the biosynthetic pathway after β-oxidation of the side chain has occurred (von Reuss et al., 2012). The exogenous addition of a synthetic ascaroside with a nine-carbon side chain, asc-C9 (ascr#10), to daf-22 worms (which are defective in β-oxidation and thus cannot make short- and medium-chain ascarosides, but which presumably can still biosynthesize the IC group and attach it) leads to production of the corresponding IC-ascaroside, IC-asc-C9 (icas#10) (von Reuss et al., 2012). This result suggests that attachment of the IC group occurs after side-chain β-oxidation, at least in the biosynthetic pathway to IC-asc-C9. The acyl-CoA synthetase gene, acs-7, has been shown to be required for biosynthesis of the short-chain IC- and OS-modified ascarosides, IC-asc-C5 and OS-asc-C5 (osas#9), but not for medium-chain IC- and OS-ascarosides, such as IC-asc-C9 and OS-asc-C9 (osas#10) (Panda et al., 2017). Panda et al. proposed that ACS-7 activates indole-3-carboxylic acid (ICA) as the corresponding CoA-thioester (IC-CoA) and is involved in the attachment of the IC group specifically to the short-chain ascaroside asc-C5 (ascr#9) to make IC-asc-C5 (Figure 1A). However, while Panda et al. were able to show that ACS-7 activates ICA as an AMP-ester (IC-AMP), they could not show that ACS-7 catalyzes the formation of IC-CoA or attachment of the IC group to an ascaroside (Figure 1A). Their interpretation of these data was that some other protein or factor was required for the full activity of ACS-7. Panda et al. showed that mutant worms lacking lysosome-related organelles (LROs) did not make 4’-modified ascarosides, and they reported that ACS-7 was expressed in the lysosome (Panda et al., 2017). Thus, they concluded that ACS-7 was functioning inside the lysosomes in IC-ascaroside biosynthesis.

Figure 1

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Here, we show that the role of the acyl-CoA synthetase ACS-7 is not to attach the IC group to ascarosides as proposed by Panda et al., but to activate the side chains of medium-chain IC-ascarosides for shortening through β-oxidation (Figure 1B). Thus, in contrast to the previously proposed model, we show that β-oxidation of the side chain occurs after attachment of the IC head group in the biosynthesis of the short-chain IC-ascarosides. Our data show that under favorable, growth-promoting conditions, the IC group is preferentially attached to ascarosides with 8–11-carbon side chains to form aggregation pheromones. However, if conditions decline, such as during starvation, the side chains of these IC-ascarosides can be activated by ACS-7 and then shortened through peroxisomal β-oxidation cycles involving ACOX-1.1 and ACOX-3 to make the potent dauer pheromone IC-asc-C5. Consistent with its function in activating ascarosides for peroxisomal β-oxidation, ACS-7 is localized to the peroxisome, rather than the lysosome, as previously reported by Panda et al. Our results uncover the mechanism by which C. elegans responds to declining environmental conditions and converts aggregation-inducing, medium-chain IC-ascarosides to dauer-inducing, short-chain IC-ascarosides. This mechanism is likely also used by C. elegans in the biosynthesis of short-chain OS-ascarosides, which induce nematode dispersal under unfavorable conditions. Using this mechanism, C. elegans can efficiently alter the function of existing ascarosides simply by tailoring the length of the side chain, providing a novel strategy to rapidly modulate chemical signaling in response to environmental conditions.

Our data show that C. elegans can engage in dynamic tailoring of the ascarosides that it has produced. This dynamic tailoring enables the worm to respond rapidly to changing environmental conditions and modulate the nature of its chemical message without having to synthesize new ascarosides de novo. Specifically, we have shown that C. elegans attaches the IC head group to ascarosides with medium-length (8–11) side chains to generate aggregation pheromones such as IC-asc-ΔC9 (icas#3) (Srinivasan et al., 2012). Production of aggregation pheromones likely occurs under favorable environmental conditions such as food-rich conditions as C. elegans tends to aggregate on food (Srinivasan et al., 2012). We have shown that C. elegans can activate IC-ascarosides with medium-length side chains for shortening of the side chains through β-oxidation to generate the dauer pheromone IC-asc-C5 (icas#9) (Figure 1B). This β-oxidation process likely occurs when environmental conditions become less favorable, such as during starvation. Indeed, starvation conditions have been shown to lead to an increase in the short-chain IC-ascaroside IC-asc-C5 in L1 larvae (Artyukhin et al., 2013). Consistent with this result, bacterial food has been shown to downregulate production of IC-asc-C5 in L4 larvae (Zhang et al., 2015). Increased production of short-chain IC-ascarosides by starvation conditions would be consistent with their biological role as IC-asc-C5 is a dauer pheromone (Butcher et al., 2009a), and C. elegans would likely want to enter dauer under starvation conditions. Our work has thus determined the mechanism by which C. elegans modulates the balance between aggregation-inducing, medium-chain IC-ascarosides and dauer-inducing, short-chain IC-ascarosides in response to changing environmental conditions.

Our model for biosynthesis of the short-chain IC-ascarosides, in which β-oxidation of the ascaroside side chain occurs after attachment of the IC head group, contrasts with the model proposed by Panda et al., in which β-oxidation of the ascaroside side chain occurs before attachment of the IC head group (Panda et al., 2017) (Figure 1A,B). We have shown that the acyl-CoA synthetase ACS-7 activates the side chains of medium-chain IC-ascarosides, such as IC-asc-C9, as CoA-thioesters, thereby enabling the side chains to be shortened through β-oxidation (Figure 1B). Previously, Panda et al. proposed that ACS-7 activates ICA as the corresponding CoA-thioester, IC-CoA, and enables attachment of the IC group to asc-C5 to make IC-asc-C5 (Panda et al., 2017) (Figure 1A). However, ACS-7 could only be shown to convert ICA to IC-AMP (Panda et al., 2017). Our data show that IC-asc-C9 is a much better substrate for ACS-7, which rapidly converts IC-asc-C9 to the corresponding CoA-thioester, IC-asc-C9-CoA (Figure 1B). We have shown that once the side chain of the medium-chain IC-ascarosides is activated, it can be shortened through β-oxidation cycles involving ACOX-1.1, ACOX-3, and likely ACOX-1.4 as well, to make the short-chain IC-ascaroside, IC-asc-C5. Specifically, we have shown that the acox-1.1;acox-3 double deletion mutant fails to make any IC-asc-C5 and accumulates IC-asc-C7. Furthermore, ACOX-1.1 and ACOX-3 are able to process IC-asc-C9-CoA to IC-asc-C5-CoA in vitro with the help of the β-oxidation enzymes MAOC-1, DHS-28, and DAF-22. Consistent with ACS-7 working in the same biosynthetic pathway as ACOX-1.1 and ACOX-3, the three genes appear to be transcriptionally co-regulated (von Mering et al., 2005). ACOX-3 has been shown to be induced by dietary restriction (Palgunow et al., 2012). ACS-7 and ACOX-3 have also been shown to be co-regulated by the transcription factor SKN-1, which functions in a variety of stress responses, including detoxification, pathogen defense, the unfolded protein response, and lipid metabolism under starvation conditions (Blackwell et al., 2015). However, the significance of this regulation for IC-ascaroside biosynthesis remains to be seen.

In addition to the IC-ascarosides, ACS-7 is also necessary for biosynthesis of short-chain OS-ascarosides, as acs-7 mutant worms cannot make the short-chain OS-ascarosides (Panda et al., 2017). Thus, it is likely that, in addition to medium-chain IC-ascarosides, ACS-7 also activates medium-chain OS-ascarosides for β-oxidation of their side chains. The β-oxidation of IC- and OS-modified ascarosides may be regulated in a similar fashion. Starvation has been shown to increase the ratio of short-chain (C5) to medium-chain (C9) OS-ascarosides (Artyukhin et al., 2013). Production of the short-chain OS-ascaroside OS-asc-C5 (osas#9) under starvation conditions is consistent with its biological role, as it acts as a dispersal signal that encourages the worm population to seek more favorable conditions (Artyukhin et al., 2013). In addition to the IC and OS head groups, the 4’-position of the ascarosides can also be modified by the MB and HB head groups (von Reuss et al., 2012). However, MB- and HB-ascarosides with fewer than nine-carbons in their side chains have not been detected in C. elegans (von Reuss et al., 2012). Thus, unlike medium-chain IC-and OS-ascarosides, medium-chain MB- and HB-ascarosides probably do not undergo activation by an acyl-CoA synthetase for further β-oxidation in the peroxisome.

In contrast to previous reports that suggested ACS-7 is localized to the lysosomes in the intestine (Panda et al., 2017), we have shown that ACS-7 is localized to the peroxisomes. It is likely that the peroxisome is where ACS-7 contributes to IC-ascaroside biosynthesis, as ACS-7 activates the medium-chain IC-ascarosides as their CoA-thioesters for β-oxidation by the peroxisomal enzymes ACOX-1.1/ACOX-3, MAOC-1, DHS-28, and DAF-22. The lysosome has been shown to be important for biosynthesis of ascarosides modified with a head group on the 4’-position of the ascarylose sugar (Panda et al., 2017). Mutant worms that lack LROs, such as glo-1 mutants, do not make any ascarosides modified on the 4’-position with IC, OS, HB, or MB head groups (Panda et al., 2017). In a model that accounts for the role of the LROs in ascaroside biosynthesis, long-chain ascarosides could be shortened to medium- and short-chain ascarosides in the peroxisome, and then medium-chain ascarosides, such as asc-C9, may be transported from the peroxisome to the LROs for attachment of 4’-modifications, such as attachment of the IC head group to make IC-asc-C9, by unknown enzymes (Figure 1B). In response to changing internal or external conditions, these modified ascarosides could then be transported back to the peroxisome for activation by ACS-7 and further β-oxidation of their side chains (Figure 1B). Trafficking of the ascarosides through multiple cellular organelles, such as the peroxisome and LROs, during biosynthesis may enable C. elegans to induce the production of modified ascarosides with specific side-chain lengths only under certain conditions or only in certain tissues.

All data generated or analysed during this study are included in the manuscript and supporting files.

1. 1. Arora P
   2. Vats A
   3. Saxena P
   4. Mohanty D
   5. Gokhale RS

   (2005) Promiscuous fatty acyl CoA ligases produce acyl-CoA and acyl-SNAC precursors for polyketide biosynthesis

   Journal of the American Chemical Society 127:9388–9389.

   https://doi.org/10.1021/ja052991s

   • PubMed
   • Google Scholar
2. 1. Arribere JA
   2. Bell RT
   3. Fu BX
   4. Artiles KL
   5. Hartman PS
   6. Fire AZ

   (2014) Efficient marker-free recovery of custom genetic modifications with CRISPR/Cas9 in Caenorhabditis elegans

   Genetics 198:837–846.

   https://doi.org/10.1534/genetics.114.169730

   • PubMed
   • Google Scholar
3. 1. Artyukhin AB
   2. Yim JJ
   3. Srinivasan J
   4. Izrayelit Y
   5. Bose N
   6. von Reuss SH
   7. Jo Y
   8. Jordan JM
   9. Baugh LR
   10. Cheong M
   11. Sternberg PW
   12. Avery L
   13. Schroeder FC

   (2013) Succinylated octopamine ascarosides and a new pathway of biogenic amine metabolism in Caenorhabditis elegans

   Journal of Biological Chemistry 288:18778–18783.

   https://doi.org/10.1074/jbc.C113.477000

   • PubMed
   • Google Scholar
4. 1. Blackwell TK
   2. Steinbaugh MJ
   3. Hourihan JM
   4. Ewald CY
   5. Isik M

   (2015) SKN-1/Nrf, stress responses, and aging in Caenorhabditis elegans

   Free Radical Biology and Medicine 88:290–301.

   https://doi.org/10.1016/j.freeradbiomed.2015.06.008

   • PubMed
   • Google Scholar
5. 1. Butcher RA
   2. Fujita M
   3. Schroeder FC
   4. Clardy J

   (2007) Small-molecule pheromones that control dauer development in Caenorhabditis elegans

   Nature Chemical Biology 3:420–422.

   https://doi.org/10.1038/nchembio.2007.3

   • PubMed
   • Google Scholar
6. 1. Butcher RA
   2. Ragains JR
   3. Kim E
   4. Clardy J

   (2008) A potent dauer pheromone component in Caenorhabditis elegans that acts synergistically with other components

   PNAS 105:14288–14292.

   https://doi.org/10.1073/pnas.0806676105

   • PubMed
   • Google Scholar
7. 1. Butcher RA
   2. Ragains JR
   3. Clardy J

   (2009a) An indole-containing dauer pheromone component with unusual dauer inhibitory activity at higher concentrations

   Organic Letters 11:3100–3103.

   https://doi.org/10.1021/ol901011c

   • PubMed
   • Google Scholar
8. 1. Butcher RA
   2. Ragains JR
   3. Li W
   4. Ruvkun G
   5. Clardy J
   6. Mak HY

   (2009b) Biosynthesis of the Caenorhabditis elegans dauer pheromone

   PNAS 106:1875–1879.

   https://doi.org/10.1073/pnas.0810338106

   • PubMed
   • Google Scholar
9. 1. Butcher RA

   (2017) Decoding chemical communication in nematodes

   Natural Product Reports 34:472–477.

   https://doi.org/10.1039/C7NP00007C

   • PubMed
   • Google Scholar
10. 1. Cassada RC
    2. Russell RL

    (1975) The dauerlarva, a post-embryonic developmental variant of the nematode Caenorhabditis elegans

    Developmental Biology 46:326–342.

    https://doi.org/10.1016/0012-1606(75)90109-8

    • PubMed
    • Google Scholar
11. 1. Cong L
    2. Zhang F

    (2015) Genome engineering using CRISPR-Cas9 system

    Methods in Molecular Biology 1239:197–217.

    https://doi.org/10.1007/978-1-4939-1862-1_10

    • PubMed
    • Google Scholar
12. 1. Farboud B
    2. Meyer BJ

    (2015) Dramatic enhancement of genome editing by CRISPR/Cas9 through improved guide RNA design

    Genetics 199:959–971.

    https://doi.org/10.1534/genetics.115.175166

    • PubMed
    • Google Scholar
13. 1. Golden JW
    2. Riddle DL

    (1982) A pheromone influences larval development in the nematode Caenorhabditis elegans

    Science 218:578–580.

    https://doi.org/10.1126/science.6896933

    • PubMed
    • Google Scholar
14. 1. Golden JW
    2. Riddle DL

    (1985) A gene affecting production of the Caenorhabditis elegans dauer-inducing pheromone

    MGG Molecular & General Genetics 198:534–536.

    https://doi.org/10.1007/BF00332953

    • PubMed
    • Google Scholar
15. 1. Greene JS
    2. Brown M
    3. Dobosiewicz M
    4. Ishida IG
    5. Macosko EZ
    6. Zhang X
    7. Butcher RA
    8. Cline DJ
    9. McGrath PT
    10. Bargmann CI

    (2016) Balancing selection shapes density-dependent foraging behaviour

    Nature 539:254–258.

    https://doi.org/10.1038/nature19848

    • PubMed
    • Google Scholar
16. 1. Hall AM
    2. Smith AJ
    3. Bernlohr DA

    (2003) Characterization of the Acyl-CoA synthetase activity of purified murine fatty acid transport protein 1

    Journal of Biological Chemistry 278:43008–43013.

    https://doi.org/10.1074/jbc.M306575200

    • PubMed
    • Google Scholar
17. 1. Hermann GJ
    2. Schroeder LK
    3. Hieb CA
    4. Kershner AM
    5. Rabbitts BM
    6. Fonarev P
    7. Grant BD
    8. Priess JR

    (2005) Genetic analysis of lysosomal trafficking in Caenorhabditis elegans

    Molecular Biology of the Cell 16:3273–3288.

    https://doi.org/10.1091/mbc.E05-01-0060

    • PubMed
    • Google Scholar
18. 1. Hisanaga Y
    2. Ago H
    3. Nakagawa N
    4. Hamada K
    5. Ida K
    6. Yamamoto M
    7. Hori T
    8. Arii Y
    9. Sugahara M
    10. Kuramitsu S
    11. Yokoyama S
    12. Miyano M

    (2004) Structural basis of the substrate-specific two-step catalysis of long chain fatty acyl-CoA synthetase dimer

    Journal of Biological Chemistry 279:31717–31726.

    https://doi.org/10.1074/jbc.M400100200

    • PubMed
    • Google Scholar
19. 1. Hollister KA
    2. Conner ES
    3. Zhang X
    4. Spell M
    5. Bernard GM
    6. Patel P
    7. de Carvalho AC
    8. Butcher RA
    9. Ragains JR

    (2013) Ascaroside activity in Caenorhabditis elegans is highly dependent on chemical structure

    Bioorganic & Medicinal Chemistry 21:5754–5769.

    https://doi.org/10.1016/j.bmc.2013.07.018

    • PubMed
    • Google Scholar
20. 1. Izrayelit Y
    2. Srinivasan J
    3. Campbell SL
    4. Jo Y
    5. von Reuss SH
    6. Genoff MC
    7. Sternberg PW
    8. Schroeder FC

    (2012) Targeted metabolomics reveals a male pheromone and sex-specific ascaroside biosynthesis in Caenorhabditis elegans

    ACS Chemical Biology 7:1321–1325.

    https://doi.org/10.1021/cb300169c

    • PubMed
    • Google Scholar
21. 1. Joo HJ
    2. Kim KY
    3. Yim YH
    4. Jin YX
    5. Kim H
    6. Kim MY
    7. Paik YK

    (2010) Contribution of the peroxisomal acox gene to the dynamic balance of daumone production in Caenorhabditis elegans

    The Journal of Biological Chemistry 285:29319–29325.

    https://doi.org/10.1074/jbc.M110.122663

    • PubMed
    • Google Scholar
22. 1. Joo HJ
    2. Yim YH
    3. Jeong PY
    4. Jin YX
    5. Lee JE
    6. Kim H
    7. Jeong SK
    8. Chitwood DJ
    9. Paik YK

    (2009) Caenorhabditis elegans utilizes dauer pheromone biosynthesis to dispose of toxic peroxisomal fatty acids for cellular homoeostasis

    Biochemical Journal 422:61–71.

    https://doi.org/10.1042/BJ20090513

    • PubMed
    • Google Scholar
23. 1. Kim H
    2. Ishidate T
    3. Ghanta KS
    4. Seth M
    5. Conte D
    6. Shirayama M
    7. Mello CC

    (2014) A co-CRISPR strategy for efficient genome editing in Caenorhabditis elegans

    Genetics 197:1069–1080.

    https://doi.org/10.1534/genetics.114.166389

    • PubMed
    • Google Scholar
24. 1. Magnes C
    2. Sinner FM
    3. Regittnig W
    4. Pieber TR

    (2005) LC/MS/MS method for quantitative determination of long-chain fatty acyl-CoAs

    Analytical Chemistry 77:2889–2894.

    https://doi.org/10.1021/ac048314i

    • PubMed
    • Google Scholar
25. 1. Palgunow D
    2. Klapper M
    3. Döring F

    (2012) Dietary restriction during development enlarges intestinal and hypodermal lipid droplets in Caenorhabditis elegans

    PLoS One 7:e46198.

    https://doi.org/10.1371/journal.pone.0046198

    • PubMed
    • Google Scholar
26. 1. Panda O
    2. Akagi AE
    3. Artyukhin AB
    4. Judkins JC
    5. Le HH
    6. Mahanti P
    7. Cohen SM
    8. Sternberg PW
    9. Schroeder FC

    (2017) Biosynthesis of Modular Ascarosides in C. elegans

    Angewandte Chemie International Edition 56:4729–4733.

    https://doi.org/10.1002/anie.201700103

    • PubMed
    • Google Scholar
27. 1. Pungaliya C
    2. Srinivasan J
    3. Fox BW
    4. Malik RU
    5. Ludewig AH
    6. Sternberg PW
    7. Schroeder FC

    (2009) A shortcut to identifying small molecule signals that regulate behavior and development in Caenorhabditis elegans

    PNAS 106:7708–7713.

    https://doi.org/10.1073/pnas.0811918106

    • PubMed
    • Google Scholar
28. 1. Srinivasan J
    2. Kaplan F
    3. Ajredini R
    4. Zachariah C
    5. Alborn HT
    6. Teal PE
    7. Malik RU
    8. Edison AS
    9. Sternberg PW
    10. Schroeder FC

    (2008) A blend of small molecules regulates both mating and development in Caenorhabditis elegans

    Nature 454:1115–1118.

    https://doi.org/10.1038/nature07168

    • PubMed
    • Google Scholar
29. 1. Srinivasan J
    2. von Reuss SH
    3. Bose N
    4. Zaslaver A
    5. Mahanti P
    6. Ho MC
    7. O'Doherty OG
    8. Edison AS
    9. Sternberg PW
    10. Schroeder FC

    (2012) A modular library of small molecule signals regulates social behaviors in Caenorhabditis elegans

    PLoS Biology 10:e1001237.

    https://doi.org/10.1371/journal.pbio.1001237

    • PubMed
    • Google Scholar
30. 1. Tanaka T
    2. Hosaka K
    3. Hoshimaru M
    4. Numa S

    (1979) Purification and properties of long-chain acyl-coenzyme-A synthetase from rat liver

    European Journal of Biochemistry 98:165–172.

    https://doi.org/10.1111/j.1432-1033.1979.tb13173.x

    • PubMed
    • Google Scholar
31. 1. Thieringer H
    2. Moellers B
    3. Dodt G
    4. Kunau WH
    5. Driscoll M

    (2003) Modeling human peroxisome biogenesis disorders in the nematode Caenorhabditis elegans

    Journal of Cell Science 116:1797–1804.

    https://doi.org/10.1242/jcs.00380

    • PubMed
    • Google Scholar
32. 1. Van Horn CG
    2. Caviglia JM
    3. Li LO
    4. Wang S
    5. Granger DA
    6. Coleman RA

    (2005) Characterization of recombinant long-chain rat acyl-CoA synthetase isoforms 3 and 6: identification of a novel variant of isoform 6

    Biochemistry 44:1635–1642.

    https://doi.org/10.1021/bi047721l

    • PubMed
    • Google Scholar
33. 1. von Mering C
    2. Jensen LJ
    3. Snel B
    4. Hooper SD
    5. Krupp M
    6. Foglierini M
    7. Jouffre N
    8. Huynen MA
    9. Bork P

    (2005) STRING: known and predicted protein-protein associations, integrated and transferred across organisms

    Nucleic Acids Research 33:D433–D437.

    https://doi.org/10.1093/nar/gki005

    • PubMed
    • Google Scholar
34. 1. von Reuss SH
    2. Bose N
    3. Srinivasan J
    4. Yim JJ
    5. Judkins JC
    6. Sternberg PW
    7. Schroeder FC

    (2012) Comparative metabolomics reveals biogenesis of Ascarosides, a modular library of small-molecule signals in C. elegans

    Journal of the American Chemical Society 134:1817–1824.

    https://doi.org/10.1021/ja210202y

    • PubMed
    • Google Scholar
35. 1. Wanders RJ
    2. Waterham HR

    (2006) Biochemistry of mammalian peroxisomes revisited

    Annual Review of Biochemistry 75:295–332.

    https://doi.org/10.1146/annurev.biochem.74.082803.133329

    • PubMed
    • Google Scholar
36. 1. Zhang X
    2. Feng L
    3. Chinta S
    4. Singh P
    5. Wang Y
    6. Nunnery JK
    7. Butcher RA

    (2015) Acyl-CoA oxidase complexes control the chemical message produced by Caenorhabditis elegans

    PNAS 112:3955–3960.

    https://doi.org/10.1073/pnas.1423951112

    • PubMed
    • Google Scholar
37. 1. Zhang X
    2. Li K
    3. Jones RA
    4. Bruner SD
    5. Butcher RA

    (2016) Structural characterization of acyl-CoA oxidases reveals a direct link between pheromone biosynthesis and metabolic state in Caenorhabditis elegans

    PNAS 113:10055–10060.

    https://doi.org/10.1073/pnas.1608262113

    • PubMed
    • Google Scholar
38. 1. Zhang X
    2. Noguez JH
    3. Zhou Y
    4. Butcher RA

    (2013) Analysis of ascarosides from Caenorhabditis elegans using mass spectrometry and NMR spectroscopy

    Methods in Molecular Biology 1068:71–92.

    https://doi.org/10.1007/978-1-62703-619-1_6

    • PubMed
    • Google Scholar
39. 1. Zhang X
    2. Wang Y
    3. Perez DH
    4. Jones Lipinski RA
    5. Butcher RA

    (2018) Acyl-CoA oxidases fine-tune the production of ascaroside pheromones with specific side chain lengths

    ACS Chemical Biology 13:1048–1056.

    https://doi.org/10.1021/acschembio.7b01021

    • PubMed
    • Google Scholar

Author details

1. Yue Zhou

   Department of Chemistry, University of Florida, Gainesville, United States

   Contribution

   Conceptualization, Resources, Data curation, Formal analysis, Validation, Visualization, Methodology, Writing—review and editing

   Contributed equally with

   Yuting Wang

   Competing interests

   No competing interests declared

2. Yuting Wang

   Department of Chemistry, University of Florida, Gainesville, United States

   Contribution

   Conceptualization, Resources, Data curation, Formal analysis, Validation, Visualization, Methodology, Writing—review and editing

   Contributed equally with

   Yue Zhou

   Competing interests

   No competing interests declared

3. Xinxing Zhang

   Department of Chemistry, University of Florida, Gainesville, United States

   Contribution

   Conceptualization, Resources, Data curation, Formal analysis, Validation, Methodology, Writing—review and editing

   Competing interests

   No competing interests declared

4. Subhradeep Bhar

   Department of Chemistry, University of Florida, Gainesville, United States

   Contribution

   Resources, Investigation, Visualization

   Competing interests

   No competing interests declared

5. Rachel A Jones Lipinski

   Department of Chemistry, University of Florida, Gainesville, United States

   Contribution

   Resources

   Competing interests

   No competing interests declared

6. Jungsoo Han

   Department of Chemistry, University of Florida, Gainesville, United States

   Contribution

   Resources

   Competing interests

   No competing interests declared

7. Likui Feng

   Department of Chemistry, University of Florida, Gainesville, United States

   Contribution

   Conceptualization

   Competing interests

   No competing interests declared

8. Rebecca A Butcher

   Department of Chemistry, University of Florida, Gainesville, United States

   Contribution

   Conceptualization, Formal analysis, Supervision, Funding acquisition, Validation, Investigation, Visualization, Methodology, Writing—original draft, Project administration, Writing—review and editing

   For correspondence

   butcher@chem.ufl.edu

   Competing interests

   No competing interests declared

   "This ORCID iD identifies the author of this article:" 0000-0002-0925-4459

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