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A maternal-effect genetic incompatibility in Caenorhabditis elegans

28 Feb 2017-bioRxiv (bioRxiv)-pp 112524
TL;DR: The discovery of a selfish element causing a genetic incompatibility between strains of the nematode Caenorhabditis elegans is discovered and the results suggest that other essential genes identified by genetic screens may turn out to be components of selfish elements.
Abstract: Selfish genetic elements spread in natural populations and have an important role in genome evolution. We discovered a selfish element causing a genetic incompatibility between strains of the nematode Caenorhabditis elegans . The element is made up of sup-35 , a maternal-effect toxin that kills developing embryos, and pha-1 , its zygotically expressed antidote. pha-1 has long been considered essential for pharynx development based on its mutant phenotype, but this phenotype in fact arises from a loss of suppression of sup-35 toxicity. Inactive copies of the sup-35/pha-1 element show high sequence divergence from active copies, and phylogenetic reconstruction suggests that they represent ancestral stages in the evolution of the element. Our results suggest that other essential genes identified by genetic screens may turn out to be components of selfish elements.

SummaryĀ (4 min read)

Introduction

  • In what is perhaps the most extreme scenario, selfish elements can kill individuals that do not inherit them, leading to a genetic incompatibility between carriers and non-carriers (Beeman et al.
  • Their underlying genetic mechanisms have been resolved in only a few cases (Werren 2011).

Results

  • V from the standard laboratory strain N2 into the strain DL238 by performing eight rounds of backcrossing and selection.
  • The authors examined the sequences of sup-35 and pha-1 in 152 C. elegans wild isolates that represented unique isotypes (Cook et al. 2016) in the Caenorhabditis elegans Natural Diversity Resource (Cook et al. 2017).
  • This suggested that the DL238 and QX1211 haplotypes were highly divergent from the N2 reference, and that major genomic rearrangements may have occurred.

Discussion

  • The antidote, pha-1, was originally thought to be a developmental gene, in large part due to the specific pharyngeal defects observed in mutants (Schnabel and Schnabel 1990; Granato et al.
  • An attractive possibility is that this regulation evolved as an additional mechanism to cope with sup-35 toxicity, as part of an arms race between the selfish element and its host.
  • These results suggest that the origin of the sup-35/pha-1 element involved the duplication of a pre-existing gene (rmd-2) and the recruitment of a novel gene of unknown origin in the lineage leading to C. elegans.
  • Lastly, their work highlights the importance of studying natural genetic variation for understanding gene function.

Acknowledgments:

  • The authors thank members of the Kruglyak lab for their comments.
  • Funding was provided by the Howard Hughes Medical Institute and NIH grant R01 HG004321 (L.K.).
  • E.B. is supported by a Gruss-Lipper postdoctoral fellowship from the EGL foundation.
  • All authors discussed and agreed on the final version of the manuscript.
  • The authors declare no competing financial interests.

Methods

  • C. elegans strains and mutant alleles Strains were grown using standard culturing techniques, with the exception that a modified nematode growth medium (NGM) containing 1% agar and 0.7% agarose was used to prevent burrowing of wild isolates (Brenner 1974; Andersen et al. 2015).
  • All experiments were carried out at 20oC.
  • All the strains used and generated in this study are listed in Table S2.
  • Some of the strains were provided by the CGC, which is funded by the NIH Office of Research Infrastructure Programs (P40 OD010440).
  • The construction of strains carrying the peel-1/zeel-1 allele from CB4856 (niDf9) was performed by backcross following a PCR product specific to the N2 allele amplified using the following primers: FW GCAGAGGAGGCAAAGGTGACTA; RV.

AGCACGTGTAGGCAGAAGTCAT.

  • Introgression of a Chr. V genetic marker into DL238 We introgressed the fog-2 (q71) allele from the N2 background into DL238 by performing eight consecutive rounds of backcross and selection for the feminization of the germline (fog) phenotype (Schedl and Kimble 1988).the authors.the authors.
  • Since the peel-1/zeel-1 element is active in N2 but not in DL238, the authors performed the backcross using DL238 males and feminized hermaphrodites.
  • The use of DL238 males avoided the fixation of the peel-1/zeel-1 element on Chr. I because DL238 worms do not carry the PEEL-1 toxin in their sperm.
  • The fixation of the N2 haplotype on Chr. III in the introgression strain could not be caused a paternal-effect toxin.
  • The introgressed marker, fog-2(q71), is required for spermatogenesis in hermaphrodites but not in males, and the resulting strain must reproduce by outcrossing.

Short read sequencing

  • The authors extracted genomic DNA (gDNA) using the DNeasy Blood & Tissue kit .
  • The authors prepared sequencing libraries using the Nextera protocol .
  • The authors followed the standard protocol with the following exception: they performed agarose size-selection of the Nextera libraries, extracting a ~500 bp band.
  • Libraries were quantified using the Qubit HS kit and sequenced using 300bp paired-end V3 kits on a Miseq desktop sequencer at 12 pM.

Variant calling in DL238

  • Automated preprocessing, alignment and variant calling were done using bcbio-nextgen (ver. 0.9.9) (https://bcbio-nextgen.readthedocs.io/).
  • Short reads from DL238 were aligned to the WBcel235 build of the reference N2 genome.
  • Variant calling was performed with four different software packages: GATK HaplotypeCaller (McKenna et al. 2010), Platypus (Rimmer et al. 2014), Freebayes (Garrison and Marth 2012) and Varscan (Koboldt et al. 2012).
  • Variants identified by fewer than three of the callers were filtered out, resulting in 239,131 SNVs between DL238 and.

N2.

  • Genotyping of the DL238 isolate and the DL238 Chr. V introgression strain Analyses were performed using the R Project for Statistical Genetics (https://www.r-project.org/).
  • To reduce the number of spurious SNVs, the authors restricted their analysis to SNVs for which all DL238 reads supported the DL238 allele when aligning to DL238, and no reads supported the N2 allele when aligning to N2.
  • Next morning, gravid hermaphrodites were allowed to lay eggs for 4-8 hours.
  • Embryonic lethality in the progeny of mating hermaphrodites was scored similarly, but L4 hermaphrodites were transferred together with males to a new plate, and their eggs were laid and collected in the presence of these males to guarantee continuous mating.
  • Visual inspection of DL238 short read alignments in the homozygous region revealed that pha-1 and sup-35 were very likely missing in DL238.

Microscopy

  • Larvae were transferred to a 3% agarose pad and visualized under bright field using a Nikon Eclipse 90i microscope equipped with a Photometrics CoolSNAP HQ2 CCD camera.
  • Multiple sequence alignments were visualized using Geneious ver. 10 (restricted free version) (https://www.geneious.com/).
  • In contrast, even null variants in sup-36 or sup-37, which are unlinked to sup-35/pha-1, are predicted to only reduce the embryonic lethality from 25% to 18.75% if they are recessive.
  • Furthermore, in a backcross of F1 males to QX2327 hermaphrodites, all of the F2 progeny inherit at least one functioning copy of sup-36 (or sup-37) from the QX2327 parent, and the lethality isnā€™t expected to be reduced at all.
  • For these reasons, and given the sample sizes in their screening, the authors cannot rule out the presence of hypomorphic variants weakly affecting sup-35, or even strongly affecting sup-36 and sup-37, in some of the wild isolates tested.

Validation of chimeric fusion

  • To validate the large deletion leading to the fusion of sup-35 and the Y48A6C.1 pseudogene, the authors designed primers that flanked the deletion: FW-del: GATCACGTGAGACAGGAAAAG and RV- del: CCCTTCAAAAGCACACCAAC.
  • This primer pair amplified the expected 1000 bp band in the wild isolate ED3012, which carries the deletion, but not in the reference N2 (fig. S8C).
  • As a positive control, the authors amplified the pha-1 locus using primers FW-pha-1: CCGTTTTCATCACGTTGCTCGA and RV-pha-1: TGTCGCGCACTACTGAATCAGA.
  • To confirm whether the chimeric fusion sup-35/Y48A6C.1 is expressed, the authors performed reverse transcription (RT) PCR (fig. S8D).
  • Total RNA was isolated from mixed stage N2 and ED3012 populations using the RNeasy kit , and cDNA was prepared using the SuperScript III Reverse Transcriptase kit (Thermo Fisher Scientific).

TTTTTCGCTTTCCAAACTGG, RV1: GCGAGCAACTCTTTCTCGAT, RV2:

  • The FW1-RV1 primer pair amplifies exclusively the spliced cDNA of wild type sup-35, and the FW1-RV2 primer pair amplifies exclusively the spliced cDNA of the sup-35/Y48A6C.1 chimeric fusion.
  • The authors then used blastn (Altschul et al. 1990) to search for scaffolds that had homology to sup- 35, pha-1, and genes in their vicinity.
  • Moreover, the region of interest contained several repetitive elements.
  • Finally, DL238 and QX1211 Illumina short reads were re-aligned to the assembled haplotypes and visually inspected to discard errors introduced by the assemblers.

Multiple sequence alignment

  • To study the genomic architecture and evolution of the sup-35/pha-1 element, the authors recovered the sequence at the sup-35/pha-1 locus in additional Caenorhabditis species.
  • Hmt-1 is a predicted transmembrane ABC transporter, and Y47D3A.29 encodes the catalytic subunit of DNA polymerase alpha.
  • Y48A6C.4, a predicted ortholog of S. cerevisiae IPI1, is located between hmt-1 and Y47D3A.29 in all sequenced Caenorhabditis.

Phylogenetic tree

  • The coding sequence of Y48A6C.4 was recovered in the different C. elegans isolates and other Caenorhabditis species.
  • The authors then assembled the cDNA sequences of Y48A6C.4 in each isolate.
  • To that end, the authors first aligned the sequence around the start and end positions of each exon to the alternative reference using blast (Altschul et al. 1990).
  • To create a gene tree, the cDNA sequences were in-silico translated, and the protein sequences were aligned using MAFFT (Katoh and Standley 2013).
  • The general time reversible (GTR) substitution model with gamma distributed rate variation was used as it was found to be the best fit (by minimizing the BIC criterion) using jModelTest2 (Darriba et al. 2012).

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1
A maternal-effect genetic incompatibility in Caenorhabditis elegans
Eyal Ben-David
*,ā€ 
, Alejandro Burga
*,ā€ 
, Leonid Kruglyak
ā€ 
Department of Human Genetics, Department of Biological Chemistry, and Howard Hughes Medical
Institute, University of California, Los Angeles, Los Angeles, CA 90095, USA.
ā€ 
To whom correspondence should be addressed. Email: ebd@ucla.edu (E.B); aburga@mednet.ucla.edu
(A.B.); lkruglyak@mednet.ucla.edu (L.K.).
* These authors contributed equally to this work.
Selfish genetic elements spread in natural populations and have an important role in genome evolution. We
discovered a selfish element causing a genetic incompatibility between strains of the nematode
Caenorhabditis elegans. The element is made up of sup-35, a maternal-effect toxin that kills developing
embryos, and pha-1, its zygotically expressed antidote. pha-1 has long been considered essential for
pharynx development based on its mutant phenotype, but this phenotype in fact arises from a loss of
suppression of sup-35 toxicity. Inactive copies of the sup-35/pha-1 element show high sequence divergence
from active copies, and phylogenetic reconstruction suggests that they represent ancestral stages in the
evolution of the element. Our results suggest that other essential genes identified by genetic screens may
turn out to be components of selfish elements.
.CC-BY-NC-ND 4.0 International licensea
certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under
The copyright holder for this preprint (which was notthis version posted March 1, 2017. ; https://doi.org/10.1101/112524doi: bioRxiv preprint

2
Introduction
Selfish genetic elements subvert the laws of Mendelian segregation to promote their own transmission
(Dawkins 1976; Doolittle and Sapienza 1980; Orgel and Crick 1980; Werren 2011; Sinkins 2011). In what
is perhaps the most extreme scenario, selfish elements can kill individuals that do not inherit them, leading
to a genetic incompatibility between carriers and non-carriers (Beeman et al. 1992; Werren 1997, 2011;
Hurst and Werren 2001; Lorenzen et al. 2008). Selfish elements are predicted to spread in natural
populations (Hurst and Werren 2001; Werren 2011), and consequently, there is significant interest in using
synthetic forms of such elements to drive population replacement of pathogen vectors in the wild (Chen et
al. 2007; Hammond et al. 2015). However, despite the prominent role of genetic incompatibilities in
genome evolution and their promise in pathogen control, their underlying genetic mechanisms have been
resolved in only a few cases (Werren 2011). Our laboratory previously identified the only known genetic
incompatibility in the nematode Caenorhabditis elegans (Seidel et al. 2008, 2011). The incompatibility is
caused by a selfish element composed of two tightly linked genes: peel-1, a sperm-delivered toxin, and
zeel-1, a zygotically expressed antidote. In crosses between isolates that carry the element and ones that do
not, the peel-1 toxin is delivered by the sperm to all progeny, so that only embryos that inherit the element
and the zeel-1 antidote survive. An analogous element, Maternal-effect dominant embryonic arrest (Medea)
has been previously described in the beetle Tribolium; however, the underlying genes remain unknown
(Beeman et al. 1992; Lorenzen et al. 2008).
Results
A maternal-effect genetic incompatibility in C. elegans
As part of ongoing efforts to study natural genetic variation in C. elegans, we introgressed a genetic marker
located on the right arm of Chr. V from the standard laboratory strain N2 into the strain DL238 by
performing eight rounds of backcrossing and selection. DL238 is a wild strain isolated in the Manuka
Natural Reserve, Hawaii, USA, and is one of the most highly divergent C. elegans isolates identified to
.CC-BY-NC-ND 4.0 International licensea
certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under
The copyright holder for this preprint (which was notthis version posted March 1, 2017. ; https://doi.org/10.1101/112524doi: bioRxiv preprint

3
date (Andersen et al. 2012). To confirm the success of the introgression, we genotyped the resulting strain
at single-nucleotide variants (SNVs) between DL238 and N2 by whole-genome sequencing. As expected,
with the exception of a small region on the right arm of Chr. V where the marker is located, most of the
genome was homozygous for the DL238 alleles (Fig. 1A). However, to our surprise, we observed sequence
reads supporting the N2 allele at many SNVs on Chr. III, including two large regions that were homozygous
for the N2 allele despite the eight rounds of backcrossing (Fig. 1A, Fig. S1). This observation suggested
that N2 variants located on this chromosome were strongly selected during the backcrossing.
To investigate the nature of the selection, we performed a series of crosses between the N2 and DL238
strains and examined their progeny. To avoid effects of the peel-1/zeel-1 element, which is present in N2
and absent in DL238, we performed a cross between DL238 males and a near isogenic line (NIL) that lacks
the peel-1/zeel-1 element in an otherwise N2 background (hereafter, N2 peel-1
-/-
) (Seidel et al. 2011). We
observed low baseline embryonic lethality in the F
1
generation and in the parental strains (0.26% (N = 381)
for F
1
; 0.99% (N = 304) for DL238; 0.4% (N = 242) for N2 peel-1
-/-
), and we did not observe any obvious
abnormal phenotypes in the F
1
that could explain the strong selection. However, when we allowed
heterozygous F
1
hermaphrodites from this cross to self-fertilize, we observed 25.15% (N = 855) embryonic
lethality among the F
2
progeny (Fig. 1B). Similar results were obtained for F1 hermaphrodites from the
reciprocal parental cross (26.1%, N = 398). These results suggested the presence of a novel genetic
incompatibility between N2 and DL238 that causes embryonic lethality in their F
2
progeny.
The observed pattern of embryonic lethality (no lethality in the parents nor in the F
1
; 25% lethality in the
F
2
) is consistent with an interaction between the genotype of the zygote and a maternal or paternal effect
(Fig. 1C) (Seidel et al. 2008). We hypothesized that the incompatibility could stem from a cytoplasmically-
inherited toxin that kills embryos if they lack a zygotically expressed antidote, analogous to the mechanism
of the peel-1/zeel-1 element (Seidel et al. 2008, 2011). To test this model and to discriminate between
maternal and paternal effects, we crossed heterozygous F
1
DL238 x N2 peel-1
ā€“/ā€“
males and hermaphrodites
.CC-BY-NC-ND 4.0 International licensea
certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under
The copyright holder for this preprint (which was notthis version posted March 1, 2017. ; https://doi.org/10.1101/112524doi: bioRxiv preprint

4
with DL238 hermaphrodites or males, respectively (Fig. 1B, Fig. S2). We observed 48.59% (N = 389)
lethality when F
1
hermaphrodites were crossed to DL238 males, but only baseline lethality (1.17%; N =
171) in the reciprocal cross of F
1
males to DL238 hermaphrodites. 50% lethality when the F
1
parent is the
mother and no lethality when the F1 parent is the father indicates that the incompatibility is caused by
maternal-effect toxicity that is rescued by a linked zygotic antidote (Fig. S2). We tested whether the new
incompatibility was independent from the paternal-effect peel-1/zeel-1 element by crossing DL238 and N2
worms and selfing the F
1
progeny. We observed 41.37% (N = 389) embryonic lethality among the F
2
progeny, consistent with expectation for Mendelian segregation of two independent incompatibilities
(43.75%) (Fig. 1D).
pha-1 and sup-35 constitute a selfish element that underlies the incompatibility between DL238 and
N2
To identify the genes underlying the maternal-effect incompatibility between N2 and DL238, we sequenced
the genome of DL238 using Illumina short reads and aligned those reads to the N2 reference genome. We
focused our attention on the two regions on Chr. III that were completely homozygous for the N2 allele in
the introgressed strain (Fig. 1A, Fig. S1). Inspection of short read coverage revealed a large ~50 kb region
on the right arm of the chromosome with very poor and sparse alignment to the N2 reference (Chr III:
11,086,500 ā€“ 11,145,000) (Fig. 2A). This region contains ten genes and two pseudogenes in N2. We noticed
that pha-1, annotated as an essential gene in the reference genome, appeared to be completely missing in
DL238 (Fig. 2A) (Schnabel and Schnabel 1990). pha-1 was originally identified as an essential gene
required for differentiation and morphogenesis of the pharynx, the C. elegans feeding organ (Schnabel and
Schnabel 1990). But if pha-1 is essential for embryonic development and missing in DL238, then how are
DL238 worms alive? pha-1 lethality can be fully suppressed by mutations in three other genes: sup-35, sup-
36, and sup-37 (Schnabel et al. 1991). We found no coding variants in sup-36 and sup-37 which reside on
.CC-BY-NC-ND 4.0 International licensea
certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under
The copyright holder for this preprint (which was notthis version posted March 1, 2017. ; https://doi.org/10.1101/112524doi: bioRxiv preprint

5
chromosomes IV and V, respectively (Schnabel et al. 1991) (Fig. S3). However, sup-35, which is located
12.5kb upstream of pha-1, also appeared to be missing or highly divergent in DL238 (Fig. 2A, Fig. S3).
We hypothesized that sup-35 and pha-1 could constitute a selfish element responsible for the observed
incompatibility between the N2 and DL238 isolates. In our model, sup-35 encodes a maternally-deposited
toxin that kills embryos unless they express the zygotic antidote, pha-1 (Fig. 2B). N2 worms carry the sup-
35/pha-1 element, which is missing or inactive in DL238, and F
1
hermaphrodites deposit the sup-35 toxin
in all their oocytes. 25% of their F
2
self-progeny do not inherit the element and are killed because they lack
the antidote pha-1. Consistent with our model, an RNA-sequencing time-course of C. elegans
embryogenesis showed that sup-35 transcripts are maternally provided, whereas pha-1 transcripts are first
detected in the embryo at the 100-cell stage (Hashimshony et al. 2014). To test our model, we first asked
whether sup-35 was necessary for the F
2
embryonic lethality in the N2 x DL238 cross. We crossed DL238
males to N2 peel-1
-/-
hermaphrodites carrying a null sup-35(e2223) allele (Fig. 2B). This sup-35 allele was
reported to fully rescue pha-1 associated embryonic lethality (Schnabel et al. 1991). Embryonic lethality in
the F
2
dropped from 25% to baseline in this cross (1.40%, N = 576), demonstrating that sup-35 activity
underlies the incompatibility between N2 and DL238 (Fig. 2B). We next tested whether expression of pha-
1, the zygotic antidote, was sufficient to rescue the embryonic lethality. We introgressed a pha-1 multicopy
transgene into the DL238 and N2 peel-1
-/-
strains and repeated the cross. As predicted, expression of pha-1
was sufficient to reduce embryonic lethality in the F
2
to baseline (1.49%, N = 268) (Fig. 2B). Moreover,
we reasoned that if the sup-35/pha-1 element underlies the maternal incompatibility, arrested embryos from
an N2 x DL238 cross should phenocopy pha-1 mutant embryos. We collected rare L1-arrested F
2
larvae
from an N2 peel-1
-/-
x DL238 cross and observed major morphological defects in the pharynx of these
individuals, as previously reported for pha-1 mutants (Schnabel and Schnabel 1990; Polley et al. 2014)
(Fig. 2C).
.CC-BY-NC-ND 4.0 International licensea
certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under
The copyright holder for this preprint (which was notthis version posted March 1, 2017. ; https://doi.org/10.1101/112524doi: bioRxiv preprint

References
More filters
Journal Articleā€¢DOIā€¢
03 Apr 1992-Science
TL;DR: A previously unknown class of dominant, maternal-effect lethal M factors was found to be widespread in natural populations of the flour beetle, Tribolium castaneum, collected on several continents and predicted to spread in a population, even in the absence of any additional selective advantage.
Abstract: A previously unknown class of dominant, maternal-effect lethal M factors was found to be widespread in natural populations of the flour beetle, Tribolium castaneum, collected on several continents. Such factors are integrated into the host chromosomes at variable locations and show the remarkable property of self-selection by maternal-effect lethality to all hatchlings that do not inherit a copy of the factor itself. Offspring are rescued by either paternally or maternally inherited copies. The M-bearing chromosome is thereby perpetuated at the expense of its non-M homolog. M factors that map to different regions of the genome do not rescue one another's maternal-effect lethality. Factors expressing these properties are predicted to spread in a population, even in the absence of any additional selective advantage. Similar factors also occur in the related species T. confusum.

177Ā citations


"A maternal-effect genetic incompati..." refers background in this paper

  • ...An analogous element, Maternal-effect dominant embryonic arrest (Medea) has been previously described in the beetle Tribolium; however, the underlying genes remain unknown (Beeman et al. 1992; Lorenzen et al. 2008)....

    [...]

  • ...In what is perhaps the most extreme scenario, selfish elements can kill individuals that do not inherit them, leading to a genetic incompatibility between carriers and non-carriers (Beeman et al. 1992; Werren 1997, 2011;Hurst and Werren 2001; Lorenzen et al. 2008)....

    [...]

Journal Articleā€¢DOIā€¢
12 Mar 2015-Nature
TL;DR: The temporal and spatial components of gene expression spanning embryonic development for all Caenorhabditis elegans genes are determined and it is proposed that the endoderm program dates back to the origin of multicellularity, whereas the ectoderm originated as a secondary germ layer freed from ancestral feeding functions.
Abstract: Studies of gene-expression levels in embryos of Caenorhabditis elegans and of other phyla reveal the timing and location of expression of all genes and support a model in which the endoderm program dates back to the origin of multicellularity while the ectoderm originated as a secondary germ layer freed from ancestral feeding functions. The germ-layer theory ā€” which holds that all the cells and tissues of the body can be grouped into three fundamental layers ā€” goes back to the roots of developmental biology as a discipline 150 years ago. The skin and many external organs are formed of ectoderm; the guts of endoderm, and organs in the middle, such as muscles and bones, form the mesoderm. The mesoderm seems to have been the last of the three layers to have evolved, as Itai Yanai and colleagues confirm with studies on the expression of genes in the embryo of the roundworm Caenorhabditis elegans. But which came first, the ectoderm or the endoderm? Further studies on a range of animals, including the sponge Amphimedon queenslandica, which lacks clear germ layers, show that the endoderm expresses evolutionarily older genes than the ectoderm. The authors speculate that the most primitive animals consisted of what would later become endoderm, with the ectoderm differentiating as cells were freed from the primary function of feeding. The concept of germ layers has been one of the foremost organizing principles in developmental biology, classification, systematics and evolution for 150 years (refs 1, 2, 3). Of the three germ layers, the mesoderm is found in bilaterian animals but is absent in species in the phyla Cnidaria and Ctenophora, which has been taken as evidence that the mesoderm was the final germ layer to evolve1,4,5. The origin of the ectoderm and endoderm germ layers, however, remains unclear, with models supporting the antecedence of each as well as a simultaneous origin4,6,7,8,9. Here we determine the temporal and spatial components of gene expression spanning embryonic development for all Caenorhabditis elegans genes and use it to determine the evolutionary ages of the germ layers. The gene expression program of the mesoderm is induced after those of the ectoderm and endoderm, thus making it the last germ layer both to evolve and to develop. Strikingly, the C. elegans endoderm and ectoderm expression programs do not co-induce; rather the endoderm activates earlier, and this is also observed in the expression of endoderm orthologues during the embryology of the frog Xenopus tropicalis, the sea anemone Nematostella vectensis and the sponge Amphimedon queenslandica. Querying the phylogenetic ages of specifically expressed genes reveals that the endoderm comprises older genes. Taken together, we propose that the endoderm program dates back to the origin of multicellularity, whereas the ectoderm originated as a secondary germ layer freed from ancestral feeding functions.

155Ā citations


"A maternal-effect genetic incompati..." refers background or result in this paper

  • ...embryogenesis showed that sup-35 transcripts are maternally provided, whereas pha-1 transcripts are first detected in the embryo at the 100-cell stage (Hashimshony et al. 2014)....

    [...]

  • ...Consistent with our model, an RNA-sequencing time-course of C. elegans embryogenesis showed that sup-35 transcripts are maternally provided, whereas pha-1 transcripts are first detected in the embryo at the 100-cell stage (Hashimshony et al. 2014)....

    [...]

13 Oct 2011
TL;DR: A sperm-delivered toxin and an embryo-expressed antidote form a co-adapted gene complex in C. elegans that promotes its own transmission to the detriment of organisms carrying it.
Abstract: The evolutionary fate of an allele ordinarily depends on its contribution to host fitness. Occasionally, however, genetic elements arise that are able to gain a transmission advantage while simultaneously imposing a fitness cost on their hosts. We previously discovered one such element in C. elegans that gains a transmission advantage through a combination of paternal-effect killing and zygotic self-rescue. Here we demonstrate that this element is composed of a sperm-delivered toxin, peel-1, and an embryo-expressed antidote, zeel-1. peel-1 and zeel-1 are located adjacent to one another in the genome and co-occur in an insertion/deletion polymorphism. peel-1 encodes a novel four-pass transmembrane protein that is expressed in sperm and delivered to the embryo via specialized, sperm-specific vesicles. In the absence of zeel-1, sperm-delivered PEEL-1 causes lethal defects in muscle and epidermal tissue at the 2-fold stage of embryogenesis. zeel-1 is expressed transiently in the embryo and encodes a novel six-pass transmembrane domain fused to a domain with sequence similarity to zyg-11, a substrate-recognition subunit of an E3 ubiquitin ligase. zeel-1 appears to have arisen recently, during an expansion of the zyg-11 family, and the transmembrane domain of zeel-1 is required and partially sufficient for antidote activity. Although PEEL-1 and ZEEL-1 normally function in embryos, these proteins can act at other stages as well. When expressed ectopically in adults, PEEL-1 kills a variety of cell types, and ectopic expression of ZEEL-1 rescues these effects. Our results demonstrate that the tight physical linkage between two novel transmembrane proteins has facilitated their co-evolution into an element capable of promoting its own transmission to the detriment of organisms carrying it.

137Ā citations

Journal Articleā€¢DOIā€¢
TL;DR: In this paper, a sperm-delivered toxin, peel-1, and an embryo-expressed antidote, zeel-1 are shown to gain a transmission advantage through a combination of paternal-effect killing and zygotic self-rescue.
Abstract: The evolutionary fate of an allele ordinarily depends on its contribution to host fitness. Occasionally, however, genetic elements arise that are able to gain a transmission advantage while simultaneously imposing a fitness cost on their hosts. We previously discovered one such element in C. elegans that gains a transmission advantage through a combination of paternal-effect killing and zygotic self-rescue. Here we demonstrate that this element is composed of a sperm-delivered toxin, peel-1, and an embryo-expressed antidote, zeel-1. peel-1 and zeel-1 are located adjacent to one another in the genome and co-occur in an insertion/deletion polymorphism. peel-1 encodes a novel four-pass transmembrane protein that is expressed in sperm and delivered to the embryo via specialized, sperm-specific vesicles. In the absence of zeel-1, sperm-delivered PEEL-1 causes lethal defects in muscle and epidermal tissue at the 2-fold stage of embryogenesis. zeel-1 is expressed transiently in the embryo and encodes a novel six-pass transmembrane domain fused to a domain with sequence similarity to zyg-11, a substrate-recognition subunit of an E3 ubiquitin ligase. zeel-1 appears to have arisen recently, during an expansion of the zyg-11 family, and the transmembrane domain of zeel-1 is required and partially sufficient for antidote activity. Although PEEL-1 and ZEEL-1 normally function in embryos, these proteins can act at other stages as well. When expressed ectopically in adults, PEEL-1 kills a variety of cell types, and ectopic expression of ZEEL-1 rescues these effects. Our results demonstrate that the tight physical linkage between two novel transmembrane proteins has facilitated their co-evolution into an element capable of promoting its own transmission to the detriment of organisms carrying it.

137Ā citations

Journal Articleā€¢DOIā€¢
01 Sep 2016-Genetics
TL;DR: It is found that telomere-length variation does not correlate with offspring production or longevity in C. elegans wild isolates, suggesting that naturally long telomeres play a limited role in modifying fitness phenotypes in Cournorhabditis elegans.
Abstract: Telomeres are involved in the maintenance of chromosomes and the prevention of genome instability. Despite this central importance, significant variation in telomere length has been observed in a variety of organisms. The genetic determinants of telomere-length variation and their effects on organismal fitness are largely unexplored. Here, we describe natural variation in telomere length across the Caenorhabditis elegans species. We identify a large-effect variant that contributes to differences in telomere length. The variant alters the conserved oligonucleotide/oligosaccharide-binding fold of protection of telomeres 2 (POT-2), a homolog of a human telomere-capping shelterin complex subunit. Mutations within this domain likely reduce the ability of POT-2 to bind telomeric DNA, thereby increasing telomere length. We find that telomere-length variation does not correlate with offspring production or longevity in C. elegans wild isolates, suggesting that naturally long telomeres play a limited role in modifying fitness phenotypes in C. elegans.

117Ā citations