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ATCC nars1 variants

Localization and conservation of <t>NARS1</t> variants (A) NARS1 functional domains are indicated in gray (unique N-terminal extension), beige (anticodon binding domain), and blue (catalytic domain). The position of each NARS1 variant is displayed across the top. (B) Phenotypes associated with each NARS1 variant are provided. Green indicates a PNS-only phenotype while blue indicates PNS and CNS features. (C) The conservation of affected amino acid residues. The position of each variant is indicated alongside neighboring NARS1 amino acid residues from evolutionarily diverse species. The position of the affected residue is indicated by bold, red text. Green shading indicates a PNS-only phenotype while blue shading indicates PNS and CNS features.

Nars1 Variants, supplied by ATCC, used in various techniques. Bioz Stars score: 94/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "Asparaginyl-tRNA synthetase ( NARS1 ) variants implicated in dominant neurological phenotypes display dominant-negative properties"

Article Title: Asparaginyl-tRNA synthetase ( NARS1 ) variants implicated in dominant neurological phenotypes display dominant-negative properties

Journal: Human Genetics and Genomics Advances

doi: 10.1016/j.xhgg.2025.100519

Localization and conservation of NARS1 variants (A) NARS1 functional domains are indicated in gray (unique N-terminal extension), beige (anticodon binding domain), and blue (catalytic domain). The position of each NARS1 variant is displayed across the top. (B) Phenotypes associated with each NARS1 variant are provided. Green indicates a PNS-only phenotype while blue indicates PNS and CNS features. (C) The conservation of affected amino acid residues. The position of each variant is indicated alongside neighboring NARS1 amino acid residues from evolutionarily diverse species. The position of the affected residue is indicated by bold, red text. Green shading indicates a PNS-only phenotype while blue shading indicates PNS and CNS features.

Figure Legend Snippet: Localization and conservation of NARS1 variants (A) NARS1 functional domains are indicated in gray (unique N-terminal extension), beige (anticodon binding domain), and blue (catalytic domain). The position of each NARS1 variant is displayed across the top. (B) Phenotypes associated with each NARS1 variant are provided. Green indicates a PNS-only phenotype while blue indicates PNS and CNS features. (C) The conservation of affected amino acid residues. The position of each variant is indicated alongside neighboring NARS1 amino acid residues from evolutionarily diverse species. The position of the affected residue is indicated by bold, red text. Green shading indicates a PNS-only phenotype while blue shading indicates PNS and CNS features.

Techniques Used: Functional Assay, Binding Assay, Variant Assay, Residue

NARS1 variants do not ablate protein expression or dimerization (A) Yeast protein lysates were subjected to western blot assays to detect human NARS1 proteins expressed from wild-type and mutant constructs, as indicated along the top. (B) Quantification of NARS1 protein expression. Green corresponds to variants associated with peripheral neuropathy and blue corresponds to variants associated with a peripheral and central nervous system phenotype. One-way ANOVA with Dunnett’s multiple comparison testing was used to determine statistical significance. (C) NanoLuc luciferase activity was expressed relative to firefly luciferase activity and then normalized to wild-type/wild-type activity. HEK293T cells expressing wild-type NARS1 C-LgBiT and HaloTag-SmBiT, as well as wild-type NARS1 C-LgBiT and wild-type NARS1 N-SmBiT independently, were used as controls. The normalized ratio is depicted on the y axis. Green corresponds to variants associated with peripheral neuropathy and blue corresponds to variants associated with a peripheral and central nervous system phenotype. One-way ANOVA with Dunnett’s multiple comparison testing was used to determine statistical significance. ∗ p = 0.015, ∗∗∗∗ p < 0.0001.

Figure Legend Snippet: NARS1 variants do not ablate protein expression or dimerization (A) Yeast protein lysates were subjected to western blot assays to detect human NARS1 proteins expressed from wild-type and mutant constructs, as indicated along the top. (B) Quantification of NARS1 protein expression. Green corresponds to variants associated with peripheral neuropathy and blue corresponds to variants associated with a peripheral and central nervous system phenotype. One-way ANOVA with Dunnett’s multiple comparison testing was used to determine statistical significance. (C) NanoLuc luciferase activity was expressed relative to firefly luciferase activity and then normalized to wild-type/wild-type activity. HEK293T cells expressing wild-type NARS1 C-LgBiT and HaloTag-SmBiT, as well as wild-type NARS1 C-LgBiT and wild-type NARS1 N-SmBiT independently, were used as controls. The normalized ratio is depicted on the y axis. Green corresponds to variants associated with peripheral neuropathy and blue corresponds to variants associated with a peripheral and central nervous system phenotype. One-way ANOVA with Dunnett’s multiple comparison testing was used to determine statistical significance. ∗ p = 0.015, ∗∗∗∗ p < 0.0001.

Techniques Used: Expressing, Western Blot, Mutagenesis, Construct, Comparison, Luciferase, Activity Assay

NARS1 variants are associated with reduced yeast cell growth (A) Yeast containing a doxycycline-repressible element upstream of the endogenous DED81 locus (the yeast ortholog of NARS1 ) were co-transformed with an empty, low-copy p413 vector and either wild-type or mutant NARS1 in a high-copy pAG425 vector. (B) Resulting cultures were plated on galactose and raffinose media lacking leucine and histidine, and including 10 μg/mL of doxycycline. NARS1 variants are indicated across the top and serial dilutions are noted on the left. Images were taken after 3 days of growth (top) and after 5 days of growth (bottom).

Figure Legend Snippet: NARS1 variants are associated with reduced yeast cell growth (A) Yeast containing a doxycycline-repressible element upstream of the endogenous DED81 locus (the yeast ortholog of NARS1 ) were co-transformed with an empty, low-copy p413 vector and either wild-type or mutant NARS1 in a high-copy pAG425 vector. (B) Resulting cultures were plated on galactose and raffinose media lacking leucine and histidine, and including 10 μg/mL of doxycycline. NARS1 variants are indicated across the top and serial dilutions are noted on the left. Images were taken after 3 days of growth (top) and after 5 days of growth (bottom).

Techniques Used: Transformation Assay, Plasmid Preparation, Mutagenesis

Pathogenic NARS1 variants cause dominant growth defects in yeast when co-expressed with wild-type NARS1 (A) Yeast containing a doxycycline-repressible element upstream of DED81 (the yeast ortholog of NARS1 ) were co-transformed with wild-type NARS1 in the p413 vector and either wild-type or mutant NARS1 in the pAG425 vector. (B) Resulting cultures were plated on galactose and raffinose media lacking leucine and histidine, with 10 μg/mL of doxycycline. NARS1 variants are indicated across the top and serial dilutions are noted on the left. Images were taken after 3 days of growth (top) and after 5 days of growth (bottom). (C) Yeast spot intensity at the 1:100 dilution was measured using ImageJ analysis and then normalized to the “WT + Null” sample. Error bars indicate the standard deviation. To test for statistically significant differences in yeast growth between each strain and the WT + Null strain, a one-way ANOVA with Dunnett’s multiple comparisons test was used. ∗∗ p = 0.002, ∗∗∗∗ p < 0.001. (D) Normalized mean gray density as described in (C). To test for statistically significant differences among all strains, a one-way ANOVA with Tukey’s multiple comparisons test was used. Given that G509S does not exert a dominant-negative effect, samples were not compared with this allele (indicated by a striped blue bar). ∗ p = 0.01, ∗∗ p = 0.007, ∗∗∗ p = 0.0001.

Figure Legend Snippet: Pathogenic NARS1 variants cause dominant growth defects in yeast when co-expressed with wild-type NARS1 (A) Yeast containing a doxycycline-repressible element upstream of DED81 (the yeast ortholog of NARS1 ) were co-transformed with wild-type NARS1 in the p413 vector and either wild-type or mutant NARS1 in the pAG425 vector. (B) Resulting cultures were plated on galactose and raffinose media lacking leucine and histidine, with 10 μg/mL of doxycycline. NARS1 variants are indicated across the top and serial dilutions are noted on the left. Images were taken after 3 days of growth (top) and after 5 days of growth (bottom). (C) Yeast spot intensity at the 1:100 dilution was measured using ImageJ analysis and then normalized to the “WT + Null” sample. Error bars indicate the standard deviation. To test for statistically significant differences in yeast growth between each strain and the WT + Null strain, a one-way ANOVA with Dunnett’s multiple comparisons test was used. ∗∗ p = 0.002, ∗∗∗∗ p < 0.001. (D) Normalized mean gray density as described in (C). To test for statistically significant differences among all strains, a one-way ANOVA with Tukey’s multiple comparisons test was used. Given that G509S does not exert a dominant-negative effect, samples were not compared with this allele (indicated by a striped blue bar). ∗ p = 0.01, ∗∗ p = 0.007, ∗∗∗ p = 0.0001.

Techniques Used: Transformation Assay, Plasmid Preparation, Mutagenesis, Standard Deviation, Dominant Negative Mutation

DED81 rescues the dominant growth phenotype caused by pathogenic NARS1 variants (A) Yeast containing a doxycycline-repressible element upstream of DED81 (the yeast ortholog of NARS1 ) were co-transformed with wild-type NARS1 in the p413 vector and either wild-type or mutant NARS1 in the pAG425 vector. (B) Resulting cultures were plated on galactose and raffinose media lacking leucine and histidine, and with no doxycycline. NARS1 variants are indicated along the top and serial dilutions are noted on the left. Images were taken after 3 days of growth (top) and after 5 days of growth (bottom). (C) Yeast spot intensity at the 1:100 dilution was measured using ImageJ analysis and then normalized to the “WT + Null” sample. Error bars indicate the standard deviation. To test for statistically significant differences in yeast growth between each strain and the WT + Null strain, a one-way ANOVA with Dunnett’s multiple comparisons test was used. ∗ p = 0.0356, ∗∗ p = 0.0018.

Figure Legend Snippet: DED81 rescues the dominant growth phenotype caused by pathogenic NARS1 variants (A) Yeast containing a doxycycline-repressible element upstream of DED81 (the yeast ortholog of NARS1 ) were co-transformed with wild-type NARS1 in the p413 vector and either wild-type or mutant NARS1 in the pAG425 vector. (B) Resulting cultures were plated on galactose and raffinose media lacking leucine and histidine, and with no doxycycline. NARS1 variants are indicated along the top and serial dilutions are noted on the left. Images were taken after 3 days of growth (top) and after 5 days of growth (bottom). (C) Yeast spot intensity at the 1:100 dilution was measured using ImageJ analysis and then normalized to the “WT + Null” sample. Error bars indicate the standard deviation. To test for statistically significant differences in yeast growth between each strain and the WT + Null strain, a one-way ANOVA with Dunnett’s multiple comparisons test was used. ∗ p = 0.0356, ∗∗ p = 0.0018.

Techniques Used: Transformation Assay, Plasmid Preparation, Mutagenesis, Standard Deviation

NARS1 variants retain dominant growth defects when expressed from a low-copy number vector (A) Yeast containing a doxycycline-repressible element upstream of DED81 (the yeast ortholog of NARS1 ) harboring wild-type NARS1 in the p413 vector were co-transformed with either wild-type or mutant NARS1 in the pAG415 vector. (B) Resulting cultures were plated on galactose and raffinose media lacking leucine and histidine, and containing 10 μg/mL of doxycycline. NARS1 variants are indicated along the top and serial dilutions are noted on the left. Images were taken after 3 days of growth. (C) Yeast spot intensity at the 1:100 dilution was measured using ImageJ analysis and then normalized to the “WT + Null” sample. Error bars indicate the standard deviation. To test for statistically significant differences in yeast growth between each strain and the WT + Null strain, a one-way ANOVA with Dunnett’s multiple comparisons test was used. ∗∗∗∗ p < 0.001.

Figure Legend Snippet: NARS1 variants retain dominant growth defects when expressed from a low-copy number vector (A) Yeast containing a doxycycline-repressible element upstream of DED81 (the yeast ortholog of NARS1 ) harboring wild-type NARS1 in the p413 vector were co-transformed with either wild-type or mutant NARS1 in the pAG415 vector. (B) Resulting cultures were plated on galactose and raffinose media lacking leucine and histidine, and containing 10 μg/mL of doxycycline. NARS1 variants are indicated along the top and serial dilutions are noted on the left. Images were taken after 3 days of growth. (C) Yeast spot intensity at the 1:100 dilution was measured using ImageJ analysis and then normalized to the “WT + Null” sample. Error bars indicate the standard deviation. To test for statistically significant differences in yeast growth between each strain and the WT + Null strain, a one-way ANOVA with Dunnett’s multiple comparisons test was used. ∗∗∗∗ p < 0.001.

Techniques Used: Low Copy Number, Plasmid Preparation, Transformation Assay, Mutagenesis, Standard Deviation

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Journal: Human Genetics and Genomics Advances

Article Title: Asparaginyl-tRNA synthetase ( NARS1 ) variants implicated in dominant neurological phenotypes display dominant-negative properties

doi: 10.1016/j.xhgg.2025.100519

Figure Lengend Snippet: Localization and conservation of NARS1 variants (A) NARS1 functional domains are indicated in gray (unique N-terminal extension), beige (anticodon binding domain), and blue (catalytic domain). The position of each NARS1 variant is displayed across the top. (B) Phenotypes associated with each NARS1 variant are provided. Green indicates a PNS-only phenotype while blue indicates PNS and CNS features. (C) The conservation of affected amino acid residues. The position of each variant is indicated alongside neighboring NARS1 amino acid residues from evolutionarily diverse species. The position of the affected residue is indicated by bold, red text. Green shading indicates a PNS-only phenotype while blue shading indicates PNS and CNS features.

Article Snippet: To evaluate the function of NARS1 variants, a p413 vector (ATCC, no. 87370) with no NARS1 insert (“Empty”) was transformed into the ptetO7- DED81 strain.

Techniques: Functional Assay, Binding Assay, Variant Assay, Residue

Journal: Human Genetics and Genomics Advances

Article Title: Asparaginyl-tRNA synthetase ( NARS1 ) variants implicated in dominant neurological phenotypes display dominant-negative properties

doi: 10.1016/j.xhgg.2025.100519

Figure Lengend Snippet: NARS1 variants do not ablate protein expression or dimerization (A) Yeast protein lysates were subjected to western blot assays to detect human NARS1 proteins expressed from wild-type and mutant constructs, as indicated along the top. (B) Quantification of NARS1 protein expression. Green corresponds to variants associated with peripheral neuropathy and blue corresponds to variants associated with a peripheral and central nervous system phenotype. One-way ANOVA with Dunnett’s multiple comparison testing was used to determine statistical significance. (C) NanoLuc luciferase activity was expressed relative to firefly luciferase activity and then normalized to wild-type/wild-type activity. HEK293T cells expressing wild-type NARS1 C-LgBiT and HaloTag-SmBiT, as well as wild-type NARS1 C-LgBiT and wild-type NARS1 N-SmBiT independently, were used as controls. The normalized ratio is depicted on the y axis. Green corresponds to variants associated with peripheral neuropathy and blue corresponds to variants associated with a peripheral and central nervous system phenotype. One-way ANOVA with Dunnett’s multiple comparison testing was used to determine statistical significance. ∗ p = 0.015, ∗∗∗∗ p < 0.0001.

Article Snippet: To evaluate the function of NARS1 variants, a p413 vector (ATCC, no. 87370) with no NARS1 insert (“Empty”) was transformed into the ptetO7- DED81 strain.

Techniques: Expressing, Western Blot, Mutagenesis, Construct, Comparison, Luciferase, Activity Assay

Journal: Human Genetics and Genomics Advances

Article Title: Asparaginyl-tRNA synthetase ( NARS1 ) variants implicated in dominant neurological phenotypes display dominant-negative properties

doi: 10.1016/j.xhgg.2025.100519

Figure Lengend Snippet: NARS1 variants are associated with reduced yeast cell growth (A) Yeast containing a doxycycline-repressible element upstream of the endogenous DED81 locus (the yeast ortholog of NARS1 ) were co-transformed with an empty, low-copy p413 vector and either wild-type or mutant NARS1 in a high-copy pAG425 vector. (B) Resulting cultures were plated on galactose and raffinose media lacking leucine and histidine, and including 10 μg/mL of doxycycline. NARS1 variants are indicated across the top and serial dilutions are noted on the left. Images were taken after 3 days of growth (top) and after 5 days of growth (bottom).

Article Snippet: To evaluate the function of NARS1 variants, a p413 vector (ATCC, no. 87370) with no NARS1 insert (“Empty”) was transformed into the ptetO7- DED81 strain.

Techniques: Transformation Assay, Plasmid Preparation, Mutagenesis

Journal: Human Genetics and Genomics Advances

Article Title: Asparaginyl-tRNA synthetase ( NARS1 ) variants implicated in dominant neurological phenotypes display dominant-negative properties

doi: 10.1016/j.xhgg.2025.100519

Figure Lengend Snippet: Pathogenic NARS1 variants cause dominant growth defects in yeast when co-expressed with wild-type NARS1 (A) Yeast containing a doxycycline-repressible element upstream of DED81 (the yeast ortholog of NARS1 ) were co-transformed with wild-type NARS1 in the p413 vector and either wild-type or mutant NARS1 in the pAG425 vector. (B) Resulting cultures were plated on galactose and raffinose media lacking leucine and histidine, with 10 μg/mL of doxycycline. NARS1 variants are indicated across the top and serial dilutions are noted on the left. Images were taken after 3 days of growth (top) and after 5 days of growth (bottom). (C) Yeast spot intensity at the 1:100 dilution was measured using ImageJ analysis and then normalized to the “WT + Null” sample. Error bars indicate the standard deviation. To test for statistically significant differences in yeast growth between each strain and the WT + Null strain, a one-way ANOVA with Dunnett’s multiple comparisons test was used. ∗∗ p = 0.002, ∗∗∗∗ p < 0.001. (D) Normalized mean gray density as described in (C). To test for statistically significant differences among all strains, a one-way ANOVA with Tukey’s multiple comparisons test was used. Given that G509S does not exert a dominant-negative effect, samples were not compared with this allele (indicated by a striped blue bar). ∗ p = 0.01, ∗∗ p = 0.007, ∗∗∗ p = 0.0001.

Article Snippet: To evaluate the function of NARS1 variants, a p413 vector (ATCC, no. 87370) with no NARS1 insert (“Empty”) was transformed into the ptetO7- DED81 strain.

Techniques: Transformation Assay, Plasmid Preparation, Mutagenesis, Standard Deviation, Dominant Negative Mutation

Journal: Human Genetics and Genomics Advances

Article Title: Asparaginyl-tRNA synthetase ( NARS1 ) variants implicated in dominant neurological phenotypes display dominant-negative properties

doi: 10.1016/j.xhgg.2025.100519

Figure Lengend Snippet: DED81 rescues the dominant growth phenotype caused by pathogenic NARS1 variants (A) Yeast containing a doxycycline-repressible element upstream of DED81 (the yeast ortholog of NARS1 ) were co-transformed with wild-type NARS1 in the p413 vector and either wild-type or mutant NARS1 in the pAG425 vector. (B) Resulting cultures were plated on galactose and raffinose media lacking leucine and histidine, and with no doxycycline. NARS1 variants are indicated along the top and serial dilutions are noted on the left. Images were taken after 3 days of growth (top) and after 5 days of growth (bottom). (C) Yeast spot intensity at the 1:100 dilution was measured using ImageJ analysis and then normalized to the “WT + Null” sample. Error bars indicate the standard deviation. To test for statistically significant differences in yeast growth between each strain and the WT + Null strain, a one-way ANOVA with Dunnett’s multiple comparisons test was used. ∗ p = 0.0356, ∗∗ p = 0.0018.

Article Snippet: To evaluate the function of NARS1 variants, a p413 vector (ATCC, no. 87370) with no NARS1 insert (“Empty”) was transformed into the ptetO7- DED81 strain.

Techniques: Transformation Assay, Plasmid Preparation, Mutagenesis, Standard Deviation

Journal: Human Genetics and Genomics Advances

Article Title: Asparaginyl-tRNA synthetase ( NARS1 ) variants implicated in dominant neurological phenotypes display dominant-negative properties

doi: 10.1016/j.xhgg.2025.100519

Figure Lengend Snippet: NARS1 variants retain dominant growth defects when expressed from a low-copy number vector (A) Yeast containing a doxycycline-repressible element upstream of DED81 (the yeast ortholog of NARS1 ) harboring wild-type NARS1 in the p413 vector were co-transformed with either wild-type or mutant NARS1 in the pAG415 vector. (B) Resulting cultures were plated on galactose and raffinose media lacking leucine and histidine, and containing 10 μg/mL of doxycycline. NARS1 variants are indicated along the top and serial dilutions are noted on the left. Images were taken after 3 days of growth. (C) Yeast spot intensity at the 1:100 dilution was measured using ImageJ analysis and then normalized to the “WT + Null” sample. Error bars indicate the standard deviation. To test for statistically significant differences in yeast growth between each strain and the WT + Null strain, a one-way ANOVA with Dunnett’s multiple comparisons test was used. ∗∗∗∗ p < 0.001.

Article Snippet: To evaluate the function of NARS1 variants, a p413 vector (ATCC, no. 87370) with no NARS1 insert (“Empty”) was transformed into the ptetO7- DED81 strain.

Techniques: Low Copy Number, Plasmid Preparation, Transformation Assay, Mutagenesis, Standard Deviation

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