WormMine: Gene WBGene00001519

• WormBase Gene ID: WBGene00001519
• Gene Name: gar-3
• Sequence Name: Y40H4A.1
• Brief Description: gar-3 encodes a G protein-linked muscarinic acetylcholine receptor; GAR-3 activity is required for regulation of membrane potential and excitation-contraction coupling in pharyngeal muscle and thus, for normal feeding behavior; in regulating pharyngeal pumping, GAR-3 likely acts upstream of GPB-2, a G-protein beta-subunit; GAR-3 also acts cell-autonomously in motor neurons to recruit the SPHK-1 sphingosine kinase to presynaptic acetylcholine release sites; a gar-3::gfp reporter fusion is expressed in the I3 pharyngeal neuron, extrapharyngeal neurons, and in pharyngeal muscles, where expression is highest in the terminal bulb.
• Organism: Caenorhabditis elegans
• Automated Description: Enables G protein-coupled acetylcholine receptor activity. Involved in several processes, including G protein-coupled acetylcholine receptor signaling pathway; positive regulation of acetylcholine secretion, neurotransmission; and positive regulation of protein localization to synapse. Located in plasma membrane. Expressed in body wall musculature; linker cell; neurons; non-striated muscle; and somatic nervous system. Human ortholog(s) of this gene implicated in several diseases, including Chagas disease; autoimmune disease of the nervous system (multiple); and lung disease (multiple). Is an ortholog of human CHRM1 (cholinergic receptor muscarinic 1); CHRM3 (cholinergic receptor muscarinic 3); and CHRM5 (cholinergic receptor muscarinic 5).
• Biotype: SO:0001217
• Genetic Position: V :6.49683 ±0.009424
• Length (nt): 14901

1 Organism

• Name: Caenorhabditis elegans
• Taxon Id: 6239

1 Synonyms

• Value: WBGene00001519

Genomics

4 Transcripts

WormMine ID	Sequence Name	Length (nt)	Chromosome Location
Transcript:Y40H4A.1a.1	Y40H4A.1a.1	2152	V: 14564819-14579719
Transcript:Y40H4A.1a.3	Y40H4A.1a.3	2093	V: 14564819-14579079
Transcript:Y40H4A.1b.1	Y40H4A.1b.1	2084	V: 14564820-14569975
Transcript:Y40H4A.1a.2	Y40H4A.1a.2	2025	V: 14564820-14569994

Other

2 CDSs

WormMine ID	Sequence Name	Length (nt)	Chromosome Location
CDS:Y40H4A.1a	Y40H4A.1a	1758	V: 14565068-14565155
CDS:Y40H4A.1b	Y40H4A.1b	1836	V: 14565068-14565155

6 RNAi Result

• WBRNAi00056336
• WBRNAi00012332
• WBRNAi00020457
• WBRNAi00027900
• WBRNAi00092909
• WBRNAi00108394

594 Allele

• gk963271
• gk962657
• gk963706
• gk962658
• gk963301
• gk964458
• gk964459
• gk963796
• ttTi43185
• gk541358
• WBVar01872555
• tm11465
• WBVar02062377
• WBVar02124257
• WBVar02062370
• WBVar02062372
• WBVar02062371
• WBVar02062374
• WBVar02062373
• WBVar02062376
• WBVar02062375
• WBVar02062378
• WBVar02062379
• WBVar02062381
• WBVar02062380
• WBVar02062383
• WBVar02062382
• WBVar02062385
• WBVar02062384
• WBVar02062387

1 Chromosome

• WormBase ID: V
• Organism: Caenorhabditis elegans
• Length (nt): 20924180

1 Chromosome Location

• Feature . Primary Identifier: WBGene00001519
• Start: 14564819
• End: 14579719
• Strand: -1

4 Data Sets

• WormBaseAcedbConverter
• GO Annotation data set
• C. elegans genomic annotations (GFF3 Gene)
• Panther orthologue and paralogue predictions

1 Downstream Intergenic Region

• WormBase ID: intergenic_region_chrV_14564640..14564818
• Length (nt): 179
• Chromosome Location: V: 14564640-14564818
• Organism: Caenorhabditis elegans

184 Expression Clusters

Regulated By Treatment	Description	Algorithm	Primary Identifier
	Transcripts that showed significantly increased expression in L1 neural cells comparing to in adult neural cells.	DESeq2 (v1.18.1) fold change > 2, P-adj<0.05, using BenjaminiHochberg correction.	WBPaper00060811:L1_vs_adult_upregulated_neural
	Transcripts expressed in neuronal cells, by analyzingfluorescence-activated cell sorted (FACS) neurons.	DESeq. False discovry rate (FDR) < 0.1.	WBPaper00048988:neuron_expressed
	Genes significantly enriched (> 2x, FDR < 5%) in a particular cell-type versus a reference sample of all cells at the same stage.	A Mann-Whitney U test with an empirical background model and FDR correction for multiple testing was used to detect expressed transcripts (Benjamini and Hochberg 1995). Genes and TARs with an FDR <= 0.05 were reported as expressed above background. Authors detected differentially expressed transcripts using a method based on linear models. Genes and TARs were called differentially expressed if the FDR was <= 0.05 and the fold change (FC) >= 2.0. To more strictly correct for potential false-positives resulting from multiple sample comparisons, authors divided individual FDR estimates by the number of samplesor sample comparisons, respectively. This resulted in an adjusted FDR of 1.3 * 0.0001 for expression above background and of 7.4 * 0.0001 for differential expression. Authors called genes selectively enriched in a given tissue if they met the following requirements: (1) enriched expression in a given tissue (FDR <= 0.05 and FC >= 2.0), (2) fold change versus reference among the upper 40% of the positive FC range observed for this gene across all tissues, and (3) fold-change entropy among the lower 40% of the distribution observed for all genes.	WBPaper00037950:A-class-motor-neurons_larva_enriched
adult vs dauer larva	Transcripts that showed differential expression in adult vs dauer lava in N2 animals at 20C.	N.A.	WBPaper00050488:adult_vs_dauer_regulated_N2_20C
Osmotic stress	Transcripts that showed significantly altered expression with 500 mM salt (NaCl) vs 100 mM salt when food was present	DESeq(version 1.10.1), FDR < 0.05.	WBPaper00050726:OsmoticStress_regulated_Food
	Neuronally enriched transcripts according to a comparison of neuronal nuclei IP samples to total nuclei using isolation of nuclei from tagged specific cell types (INTACT) technology.	DESEQ2, fold change > 2 and FDR < 0.01.	WBPaper00062103:neuron_enriched
	Transcripts that showed significantly increased expression in csr-1a(tor159) comparing to in N2 at 25C.	DESeq2, fold change > 2, p-value < 0.05.	WBPaper00061753:csr-1(tor159)_upregulated_25C
	Transcripts that showed significantly increased expression glp-1(e2141); TU3401 animals comparing to in TU3401 animals.	Fold change > 2, FDR < 0.01.	WBPaper00065993:glp-1(e2141)_upregulated
Bacteria infection: Enterococcus faecalis	Genes with increased expression after 24 hours of infection by E.faecalis Fold changes shown are pathogen vs OP50.	For RNA-seq and tiling arrays, log2 fold changes between gene expression values of infected versus uninfected nematodes were calculated. For log2 fold changes > 0.00001 the values > 81.25th percentile were defined as up-regulated and for log2 fold changes < -0.00001 the values < 18.75th percentile were defined as down-regulated.	WBPaper00038438:E.faecalis_24hr_upregulated_TilingArray
	Transcripts that showed significantly decreased expression at 5-days-post L4 adult N2 hermaphrodites comparing to 1-day-post L4 adult N2 hermaphrodites.	DESeq2, fold change > 2, FDR < 0.05	WBPaper00065835:Day5_vs_Day1_downregulated
	Transcripts expressed in GABAergic neuron, according to PAT-Seq analysis using Punc-47-GFP-3XFLAG mRNA tagging.	Cufflinks FPKM value >=1.	WBPaper00050990:GABAergic-neuron_expressed
	Genes with expression level regulated by genotype (N2 vs CB4856) and age at L3 larva and Late reproduction stage (96 hours at 24 centigrade).	For model 2, authors used 100 permutations to estimate the FDR threshold. Per permutation, genotypes and ages were independently randomly distributed, keeping the among-gene structure intact. Then for each spot (23,232) on the array, model 2 was tested. The obtained P-values were used to estimate a threshold for each of the explanatory factors. Authors also used a genome-wide threshold of -log10 P-value = 2, which resembles an FDR of 0.072 and 0.060 for marker and the interaction age-marker for the developing worms and FDR of 0.050 and 0.065 for marker and age-marker for the aging worms. For the physiological age effect, authors used a log10 P-value = 8 in developing worms (0.012 FDR) and -log10 P-value = 6 (0.032 FDR).	WBPaper00040858:eQTL_age_regulated_developing
	Transcripts that showed significantly decreased expression in day 3 adult hermaphrodite comparing to in L4 larva daf-16(mu86);glp-1(e2141) animals.	Fold change > 2, FDR < 0.05	WBPaper00064088:Day-3-adult_vs_L4_downregulated_daf-16(mu86);glp-1(e2141)
	Transcripts that showed significantly decreased expression in day 3 adult hermaphrodite comparing to in L4 larva glp-1(e2141) animals.	Fold change > 2, FDR < 0.05	WBPaper00064088:Day-3-adult_vs_L4_downregulated_glp-1(e2141)
	Transcripts that showed significantly increased expression in rrf-3(pk1426) comparing to in N2 at embryo stage.	DESeq2v 1.18.1, fold change > 1.5, adjusted p-value < 0.01.	WBPaper00056169:rrf-3(pk1426)_upregulated_embryo
	Transcripts that showed significantly increased expression after four-day-old young adult worms were placed on NGM plates seeded with OP50 in the presence 5% Agaro-oligosaccharides(AGO) for 24 h, comparing to animals grown in the absence of AGO.	Fold change > 2.	WBPaper00064306:Agaro-oligosaccharides_upregulated
	Transcripts that showed significantly increased expression in sin-3(tm1276) comparing to in N2.	DESeq2, fold change > 2, p-value < 0.01.	WBPaper00061203:sin-3(tm1276)_upregulated
	Transcripts that showed significantly increased expression in aak-1(tm1944);aak-2(ok524) animals comparing to in N2.	DEseq 1.18.0, adjusted p-value < 0.05.	WBPaper00056471:aak-1(tm1944);aak-2(ok524)_upregulated
	Transcripts that showed significantly decreased expression in N2 animals exposed to 0.1mM Paraquat from hatching to reaching adult stage.	DESeq2 version 1.22.2, p < 0.05	WBPaper00064716:paraquat_downregulated
	Transcripts that showed significantly altered expression after 24 hour exposure to stavudine (d4T) starting at L1 lava stage.	DESeq	WBPaper00053302:stavudine_24h_regulated
	Transcripts that showed significantly decreased expression in sin-3(tm1276) comparing to in N2 at early embryo when there were only 3 -5 eggs in the adult.	DESeq2, fold change > 2, adjusted p-value < 0.01	WBPaper00058598:sin-3(tm1276)_downregulated
	Genes that showed expression levels higher than the corresponding reference sample (L2 all cell reference).	A Mann-Whitney U test with an empirical background model and FDR correction for multiple testing was used to detect expressed transcripts (Benjamini and Hochberg 1995). Genes and TARs with an FDR <= 0.05 were reported as expressed above background. Authors detected differentially expressed transcripts using a method based on linear models. Genes and TARs were called differentially expressed if the FDR was <= 0.05 and the fold change (FC) >= 2.0. To more strictly correct for potential false-positives resulting from multiple sample comparisons, authors divided individual FDR estimates by the number of samplesor sample comparisons, respectively. This resulted in an adjusted FDR of 1.3 * 0.0001 for expression above background and of 7.4 * 0.0001 for differential expression. Authors called genes selectively enriched in a given tissue if they met the following requirements: (1) enriched expression in a given tissue (FDR <= 0.05 and FC >= 2.0), (2) fold change versus reference among the upper 40% of the positive FC range observed for this gene across all tissues, and (3) fold-change entropy among the lower 40% of the distribution observed for all genes.	WBPaper00037950:A-class-motor-neurons_L2-larva_expressed
	Genes that showed expression levels higher than the corresponding reference sample (L2 all cell reference).	A Mann-Whitney U test with an empirical background model and FDR correction for multiple testing was used to detect expressed transcripts (Benjamini and Hochberg 1995). Genes and TARs with an FDR <= 0.05 were reported as expressed above background. Authors detected differentially expressed transcripts using a method based on linear models. Genes and TARs were called differentially expressed if the FDR was <= 0.05 and the fold change (FC) >= 2.0. To more strictly correct for potential false-positives resulting from multiple sample comparisons, authors divided individual FDR estimates by the number of samplesor sample comparisons, respectively. This resulted in an adjusted FDR of 1.3 * 0.0001 for expression above background and of 7.4 * 0.0001 for differential expression. Authors called genes selectively enriched in a given tissue if they met the following requirements: (1) enriched expression in a given tissue (FDR <= 0.05 and FC >= 2.0), (2) fold change versus reference among the upper 40% of the positive FC range observed for this gene across all tissues, and (3) fold-change entropy among the lower 40% of the distribution observed for all genes.	WBPaper00037950:all-neurons_L2-larva_expressed
	Transcripts that showed significantly increased expression in hpk-1(pk1393) comparing to in N2 at adult day 2.	DESeq 2, fold change > 2, FDR < 0.05.	WBPaper00065581:hpk-1(pk1393)_upregulated
	Transcripts that showed significantly increased expression in hda-1(ne4752[3xFLAG-Degron-HDA-1]) in gonads dissected from 1-day old adult animals.	Salmon was used to map the mRNA-seq reads with the worm database WS268, and its output files were imported to DESeq2 in R. The differentially expressed genes were filtered by fold change more than 2 and adjusted p-value < 0.05. The scatter plots were generated by the plot function in R.	WBPaper00061479:hda-1(ne4752)_upregulated
	Genes that showed expression levels higher than the corresponding reference sample (L3/L4 all cell reference).	A Mann-Whitney U test with an empirical background model and FDR correction for multiple testing was used to detect expressed transcripts (Benjamini and Hochberg 1995). Genes and TARs with an FDR <= 0.05 were reported as expressed above background. Authors detected differentially expressed transcripts using a method based on linear models. Genes and TARs were called differentially expressed if the FDR was <= 0.05 and the fold change (FC) >= 2.0. To more strictly correct for potential false-positives resulting from multiple sample comparisons, authors divided individual FDR estimates by the number of samplesor sample comparisons, respectively. This resulted in an adjusted FDR of 1.3 * 0.0001 for expression above background and of 7.4 * 0.0001 for differential expression. Authors called genes selectively enriched in a given tissue if they met the following requirements: (1) enriched expression in a given tissue (FDR <= 0.05 and FC >= 2.0), (2) fold change versus reference among the upper 40% of the positive FC range observed for this gene across all tissues, and (3) fold-change entropy among the lower 40% of the distribution observed for all genes.	WBPaper00037950:dopaminergic-neurons_L3-L4-larva_expressed
	Transcripts that showed significantly increased expression in mep-1(ne4629[MEP-1-GFP-Degron]) in gonads dissected from 1-day old adult animals.	Salmon was used to map the mRNA-seq reads with the worm database WS268, and its output files were imported to DESeq2 in R. The differentially expressed genes were filtered by fold change more than 2 and adjusted p-value < 0.05. The scatter plots were generated by the plot function in R.	WBPaper00061479:mep-1(ne4629)_upregulated
	Genes that showed expression levels higher than the corresponding reference sample (L2 all cell reference).	A Mann-Whitney U test with an empirical background model and FDR correction for multiple testing was used to detect expressed transcripts (Benjamini and Hochberg 1995). Genes and TARs with an FDR <= 0.05 were reported as expressed above background. Authors detected differentially expressed transcripts using a method based on linear models. Genes and TARs were called differentially expressed if the FDR was <= 0.05 and the fold change (FC) >= 2.0. To more strictly correct for potential false-positives resulting from multiple sample comparisons, authors divided individual FDR estimates by the number of samplesor sample comparisons, respectively. This resulted in an adjusted FDR of 1.3 * 0.0001 for expression above background and of 7.4 * 0.0001 for differential expression. Authors called genes selectively enriched in a given tissue if they met the following requirements: (1) enriched expression in a given tissue (FDR <= 0.05 and FC >= 2.0), (2) fold change versus reference among the upper 40% of the positive FC range observed for this gene across all tissues, and (3) fold-change entropy among the lower 40% of the distribution observed for all genes.	WBPaper00037950:GABAergic-motor-neurons_L2-larva_expressed
	Genes that showed expression levels higher than the corresponding reference sample (L2 all cell reference).	A Mann-Whitney U test with an empirical background model and FDR correction for multiple testing was used to detect expressed transcripts (Benjamini and Hochberg 1995). Genes and TARs with an FDR <= 0.05 were reported as expressed above background. Authors detected differentially expressed transcripts using a method based on linear models. Genes and TARs were called differentially expressed if the FDR was <= 0.05 and the fold change (FC) >= 2.0. To more strictly correct for potential false-positives resulting from multiple sample comparisons, authors divided individual FDR estimates by the number of samplesor sample comparisons, respectively. This resulted in an adjusted FDR of 1.3 * 0.0001 for expression above background and of 7.4 * 0.0001 for differential expression. Authors called genes selectively enriched in a given tissue if they met the following requirements: (1) enriched expression in a given tissue (FDR <= 0.05 and FC >= 2.0), (2) fold change versus reference among the upper 40% of the positive FC range observed for this gene across all tissues, and (3) fold-change entropy among the lower 40% of the distribution observed for all genes.	WBPaper00037950:glr-1(+)-neurons_L2-larva_expressed
	Genes that showed expression levels higher than the corresponding reference sample (L3/L4 all cell reference).	A Mann-Whitney U test with an empirical background model and FDR correction for multiple testing was used to detect expressed transcripts (Benjamini and Hochberg 1995). Genes and TARs with an FDR <= 0.05 were reported as expressed above background. Authors detected differentially expressed transcripts using a method based on linear models. Genes and TARs were called differentially expressed if the FDR was <= 0.05 and the fold change (FC) >= 2.0. To more strictly correct for potential false-positives resulting from multiple sample comparisons, authors divided individual FDR estimates by the number of samplesor sample comparisons, respectively. This resulted in an adjusted FDR of 1.3 * 0.0001 for expression above background and of 7.4 * 0.0001 for differential expression. Authors called genes selectively enriched in a given tissue if they met the following requirements: (1) enriched expression in a given tissue (FDR <= 0.05 and FC >= 2.0), (2) fold change versus reference among the upper 40% of the positive FC range observed for this gene across all tissues, and (3) fold-change entropy among the lower 40% of the distribution observed for all genes.	WBPaper00037950:hypodermis_L3-L4-larva_expressed

12 Expression Patterns

• Primary Identifier: Expr1030914
• Pattern: Tiling arrays expression graphs

• Primary Identifier: Expr1019599
• Pattern: Developmental gene expression time-course. Raw data can be downloaded from ftp://caltech.wormbase.org/pub/wormbase/datasets-published/levin2012

• Primary Identifier: Expr8297
• Remark: Picture: Fig. 6A, B.
• Subcellular Localization: Fusion proteins expressed from the promoter constructs were found on the membranes and processes.

• Primary Identifier: Expr8296
• Remark: Picture: Fig. 3C,D.
• Pattern: Expressed in the SPC motor neurons, PCB and PCA postcloacal sensilla (p.c.s.) neurons, male spicule protractor muscles and anal depressor muscle, VD and DD ventral cord neurons, some tail and nerve ring neurons, and body wall muscles. In hermaphrodites, YFP expression is in the same set of cells, except those specific to males.

• Primary Identifier: Expr11232
• Pattern: In transgenic animals expressing Pgar-3::gfp, fluorescence was detected in body wall and pharynx muscles, as well as in motor neurons of the ventral nerve cord. GFP fluorescence was detected in all cholinergic motor neurons of the ventral nerve cord but not in GABAergic motor neurons.
• Subcellular Localization: GAR-3-GFP fluorescence was not detected in axons in single-copy transgenic animals (0 of 15 axons) and was rarely detected in axons of multicopy animals (3 of 23 axons). GAR-3-GFP was also not detected in dendrites of single-copy transgenic animals, and was occasionally detected in proximal dendrites of multicopy animals (data not shown). When axonal or dendritic GAR-3-GFP fluorescence was detected, it adopted a diffuse pattern of localization. Based on these results, the authors conclude that GAR-3 is highly enriched in cholinergic motor neuron cell bodies, where it asymmetrically distributes to the dorsal surfaces, but is likely to be very low or absent from neuronal processes. GAR-3-GFP fluorescence was observed in the ventral nerve cord and in motor neuron cell bodies in discrete patches that run parallel to the ventral nerve cord. In motor neurons, GAR-3-GFP primarily localized to narrow regions that extended longitudinally along the dorsal side of each motor neuron cell body. This pattern is suggestive of cell surface localization of GAR-3-GFP because the fluorescent regions were found on the outer edge of cell bodies marked by soluble mCherry. Similar localization patterns were observed in DA/DB motor neurons of animals expressing single-copy GAR-3-gfp (by MosSCI insertion of Punc-129::GAR-3-gfp). In body wall muscles, GAR-3-GFP fluorescence was also asymmetrically distributed at sites of contact between muscle cells, on muscle arms, and in postsynaptic regions where muscle arms contact the dorsal and ventral nerve cords. GAR-3-GFP localization was not asymmetric early in development (asymmetry ratio 1.19 +/- 0.6 in the L3 stage), but it became asymmetric starting at the L4 stage (3.25 +/- 0.34) and maintained asymmetry in adults. GAR-3-GFP on muscle membranes was visible in young animals (L3 stage) and appeared enhanced in older animals.

• Primary Identifier: Expr11145
• Remark: Strain CG308.
• Pattern: gar-3 is expressed in the linker cell.

• Primary Identifier: Expr2011946
• Pattern: Single cell embryonic expression. Only cell types with an expression fraction of greater 0.2 of the maximum expressed fraction are labeled (Full data can be downloaded from http://caltech.wormbase.org/pub/wormbase/datasets-published/packer2019/). The colors represent the broad cell class to which the cell type has been assigned. The size of the point is proportional to the log2 of the numbers of cells in the dataset of that cell type. Interactive visualizations are available as a web app (https://cello.shinyapps.io/celegans/) and can also be installed as an R package (https://github.com/qinzhu/VisCello.celegans).

• Primary Identifier: Expr3002
• Remark: Reporter gene fusion type not specified.
• Pattern: GFP expressed in the pharyngeal muscle, in pharyngeal neuron I3, and in neurons of the extrapharyngeal nervous system. Within the pharyngeal muscle, the intensity of staining is highest in the terminal bulb, lower in the isthmus, and variable in the metacorpus.

• Primary Identifier: Expr2659
• Pattern: Two RT-PCR products of expected sizes (747 and 825 bp) were observed at all developmental stages examined, indicating that the two gar-3 isoforms are expressed throughout development. The gar-3b mRNA was more abundantly expressed than gar-3a mRNA at all developmental stages. However, the relative expression ratio of the two isoforms appeared to vary between developmental stages. The gar-3a mRNA was expressed most abundantly at the first larval stage, whereas gar-3b mRNA was expressed most abundantly at embryonic and early larval stages.

• Primary Identifier: Expr2030183
• Pattern: Single cell embryonic expression. Only cell types with an expression fraction of greater 0.2 of the maximum expressed fraction are labeled (Full data can be downloaded from http://caltech.wormbase.org/pub/wormbase/datasets-published/packer2019/). The colors represent the broad cell class to which the cell type has been assigned. The size of the point is proportional to the log2 of the numbers of cells in the dataset of that cell type. Interactive visualizations are available as a web app (https://cello.shinyapps.io/celegans/) and can also be installed as an R package (https://github.com/qinzhu/VisCello.celegans).

• Primary Identifier: Expr1138
• Pattern: All developmental stages.

• Primary Identifier: Expr1159824
• Pattern: Developmental gene expression time-course. Raw data can be downloaded from ftp://caltech.wormbase.org/pub/wormbase/datasets-published/hashimshony2015

30 GO Annotation

24 Homologues

1 Locations

• Feature . Primary Identifier: WBGene00001519
• Start: 14564819
• End: 14579719
• Strand: -1

30 Ontology Annotations

0 Regulates Expr Cluster

1 Sequence

• Length: 14901

1 Sequence Ontology Term

• Name: gene

6 Strains

• WBStrain00022220
• WBStrain00022221
• WBStrain00022222
• WBStrain00035957
• WBStrain00035965
• WBStrain00052083

1 Upstream Intergenic Region

• WormBase ID: intergenic_region_chrV_14579720..14583301
• Length (nt): 3582
• Chromosome Location: V: 14579720-14583301
• Organism: Caenorhabditis elegans