2.3.1.255 A6P NAA10 variant c.16G>C has been identified in a male with intelectual disability and hypertrophic cardiomyopathy. A6P protein is highly reduced in its capacity to form the NatA complex, and displays a moderate reduction in monomeric NAA10 N-acteyltransferase function 777587 2.3.1.255 E157K NAA10 variant c.469G>A has been identified in a baby girl with cerebellar vermian hypoplasia with cystic dilatation of the 4th ventricle, a left pneumothorax and a heart murmur. Mutation does not impair NAA10-NAA15 binding and displays a moderate reduction in monomeric NAA10 N-acteyltransferase function 777587 2.3.1.255 E24A mutation in NatA, decrease in kcat, increase in Km value 736866 2.3.1.255 E24D mutation in NatA, decrease in kcat, increase in Km value 736866 2.3.1.255 E24Q mutation in NatA, decrease in kcat, increase in Km value 736866 2.3.1.255 E61A mutation in NatA, decrease in kcat, increase in Km value 736866 2.3.1.255 E62A mutation in NatA, decrease in kcat, increase in Km value 736866 2.3.1.255 F128I site-directed mutagenesis, the mutation leads an altered structure and reduced stability, and a dramatic recuction of Nt catalytic activity compared to wild-type 756675 2.3.1.255 F128L NAA10 variant c.384T>G has been identified in a boy with mixed specific developmental disorder 777587 2.3.1.255 F128L site-directed mutagenesis, the mutation leads an altered structure and reduced stability, and a dramatic recuction of Nt catalytic activity compared to wild-type 756675 2.3.1.255 H111A mutation in NatA, decrease in kcat, Km value similar to wild-type 736866 2.3.1.255 H20A mutation in NatA, decrease in kcat, increase in Km value 736866 2.3.1.255 H72A mutation in NatA, decrease in kcat, increase in Km value 736866 2.3.1.255 I72T a naturally occuring mutation NAA10 c.215T>C, the mutant phenotype shows a milder phenotypic spectrum in comparison to most of the previously described patients with NAA10 variants. The three boys have development delay, intellectual disability, and cardiac abnormalities as overlapping phenotypes. NAA10 Ile72Thr protein is destabilized, while binding to NAA15 most likely is intact. The NatA activity of NAA10 Ile72Thr appears normal while its monomeric activity is decreased. Genotype-phenotype correlations for NAA10 variants, overview 756642 2.3.1.255 K136R site-directed mutagenesis, that lacks autoacetylation, the mutant shows wild-type NAT activity 757723 2.3.1.255 K136R site-directed mutagenesis, the non-acetylated K136R mutant shows N-terminal acetyltransferase capacity as strongly as the hARD1/NAA10 wild-type, but fails to acetylate itself 757723 2.3.1.255 K29A mutation in NatA, increase in kcat, decrease in Km value 736866 2.3.1.255 K29A/Y33A mutation in NatA, decrease in kcat, Km value similar to wild-type 736866 2.3.1.255 K59A mutation in NatA, decrease in kcat, increase in Km value 736866 2.3.1.255 K59A/E61A mutation in NatA, decrease in kcat, increase in Km value 736866 2.3.1.255 K59A/E62A mutation in NatA, decrease in kcat, increase in Km value 736866 2.3.1.255 L22A mutation in NatA, decrease in kcat, increase in Km value 736866 2.3.1.255 L814P site-directed mutagenesis, the hNAA15 mutant is defective for HYPK inhibition and reduces hNatA thermostability, hNAA10 binding is not affected. The hNAA15-L814P-V5 hNatA complex shows an increased catalytic activity compared to wild-type hNatA 757754 2.3.1.255 additional information construction of Naa10 stably knocked down H1299 cell line H1299-shNaa10, cDNA microarray analysis 758388 2.3.1.255 additional information enzyme knockdown by siRNA. Generation of mutant daf-31(m655) by removal of 151 bp of promoter upstream of the ATG start codon and 242 bp of daf-31 coding region dowstream of the ATG start codon. Generation of and overexpression mutant daf-31 OE for which the full-length dar-31 genomic DNA is cloned into pGEM-T vector. Generation of mutant vncBDk by with impaired N-terminal activity -, 756675 2.3.1.255 additional information generation of an Ard1 null mutant by removal of the ARD1 coding region 756675 2.3.1.255 additional information generation of enzyme mutant by frame shift mutation causing a acetyltransferase-truncated enzymatic region, and of another mutant vnc by intron-insertion mutation 756675 2.3.1.255 additional information generation of mutant naa10-1 and of a naa15 mutant by T-DNA insertion-disrupting gene expression 756675 2.3.1.255 additional information generation of mutant naa10MO by morpholino-based knockdown 756675 2.3.1.255 additional information generation of mutants MtRimI4-158, MtRimI1-153, MtRimI4-153, MtRimIC21A, and of the final construct MtRimIC21A4-153, MtRimIC21A4-153 has almost identical enzymatic activity compared to MtRimI, indicating insignificant influence of the recombinant variations on enzymatic functions. The 2D 1H-15N heteronuclear single quantum coherence spectrum of tRimIC21A4-153 exhibits wider chemical shift dispersion and favorable peak isolation, indicating that MtRimIC21A4-153 is amendable for further structural determination. Moreover, bio-layer interferometry experiments show that MtRimIC21A4-153 possesses similar micromolar affinity to full-length MtRimI for binding the hexapeptide substrate Ala-Arg-Tyr-Phe-Arg-Arg. Structure comparison of wild-type MtRimI and mutant MtRimIC21A4-153 -, 755712 2.3.1.255 additional information knockdown of Naa10p by shRNAs, knockdown efficiencies, overview. Generation of truncated Naa10p mutants 756332 2.3.1.255 additional information mutation ard1::HIS3 is caused by inserting a Barn HI fragment containinng the HIS3 gene into the Barn HI site of plasmid YCpE18 that lies within the functional sequence of ARD1. Genes nat1/ard1 double mutant nat1-5::LEU;ard1 is generated by mating of nat1 and ard1 single mutants, the single mutants of nat1 (Naa15) and ard1 (Naa10) display identical phenotypes, no additional phenotypes are found in the double mutant. Recombinant expression of the S37P mutant of human Naa10 in a NatA-defective yeast strain, the hNaa10 expressing mutant strain shows a lack of proper complex formation with hNaa15 and reduced in vitro catalytic activity, a decrease of Nt-acetylome and an increase in the Hsp70 family proteins -, 756675 2.3.1.255 additional information silencing of mouse immortalized embryonic endothelial cells -, 758388 2.3.1.255 additional information variant c.303C>A in subunit NAA10 has been identified in a girl with developmental delay, who also displays hemihypertrophy. c.303C>A protein is completely impaired in its ability to bind NAA15 and to form an enzymatically active NatA complex 777256 2.3.1.255 additional information variants c.1009_1012delGAAA, p.Glu337Arg*5 and c.79A>G, p.R27G are associated with pediatric hypertrophic cardiomyopathy. c.1009_1012delGAAA, p.Glu337Arg*5 results in a frameshift in the transcript followed by a premature stop codon five amino acids downstream. c.79A>G, p.R27G is a missense mutation 776377 2.3.1.255 N101K variant in subunit NAA10, identified in a girl with developmental delay, who also displays hemihypertrophy. The integrity of NAA10 N101K as a monomeric acetyltransferase is intact 777256 2.3.1.255 N132A the mutant shows 4.5fold increase in Km, with no significant difference in kcat compared to the wild type enzyme 730955 2.3.1.255 P23A mutation in NatA, decrease in kcat, increase in Km value 736866 2.3.1.255 Q129P NAA10 variant c.386A>C has been identified in a female with contractures at the major joints, spasticity, and dysconjugate gaze. Mutation does not impair NAA10-NAA15 binding but and displays almost complete loss of monomeric NAA10 N-acteyltransferase function 777587 2.3.1.255 R100A the mutant shows 7fold increase in Km, with no significant difference in kcat compared to the wild type enzyme 730955 2.3.1.255 R113A mutation in NatA, strong decrease in kcat, Km value similar to wild-type 736866 2.3.1.255 R116W site-directed mutagenesis, the mutation leads to a reduction in catalytic activity for the peptide substrates EEEI and SESS by 15% compared to wild-type 756675 2.3.1.255 R79C NAA10 variant c.235C>T has been identified in several male family members with intelectual disability. Mutation does not impair NAA10-NAA15 binding and displays a moderate reduction in monomeric NAA10 N-acteyltransferase function 777587 2.3.1.255 R80A mutation in NatA, decrease in kcat, increase in Km value 736866 2.3.1.255 R82A/Y122F site-directed mutagenesis, the mutant shows highly reduced NAT activity compared to wild-type 757723 2.3.1.255 R82A/Y122F the acetyltransferase dead DN mutant of hARD1/NAA10 almost loses its NAT activity and fails to acetylate itself. The DN mutant includes two mutations R82A and Y122F, which inhibit the binding of acetyl-CoA to hARD1/NAA10 and consequently suppresses its acetyltransferase activity 757723 2.3.1.255 R83C site-directed mutagenesis, the mutation interferes with acetyl-CoA binding and leads to a 60% reduction in Nt-catalytic activity compared to wild-type 756675 2.3.1.255 R83H naturally occuring c.248G > A missense mutation, reduced enzymatic activity of monomeric NAA10-R83H. This variant is modelled to have an altered charge density in the acetyl-CoA binding region of NAA10 756298 2.3.1.255 S37P site-directed mutagenesis, the mutant Naa10 protein shows reduced catalytic activity for EEEI, DDDI, and SESS peptide substrates, and inability to combine with Naa15. The mutant hNaa10 S37P recombinantly expressed in a NatA-defective Saccharomyces cerevisiae strain lacks a proper complex formation with hNaa15 and is reduced in in vitro catalytic activity 756675 2.3.1.255 S37P the mutation is the cause of Ogden Syndrome 741085 2.3.1.255 S39P site-directed mutagenesis, the mutation does not cause a phenotype -, 756675 2.3.1.255 T105A the mutant shows 3fold increase in Km, with no significant difference in kcat compared to the wild type enzyme 730955 2.3.1.255 T406Y site-directed mutagenesis, the hNAA15-T406Y-V5 hNatA mutant complex displays a decreased catalytic activity toward the hNatA substrate SESS compared to wild-type hNatA. the hNAA15 mutant can disassociate hNAA50 from hNatA in vitro, hNAA10 binding is not affected 757754 2.3.1.255 V107F site-directed mutagenesis, the mutation leads to a reduction in catalytic activity for the peptide substrates EEEI and SESS by 95% compared to wild-type 756675 2.3.1.255 V111G a naturally occuring 332 T > G missense mutant, the mutant Naa10 has a reduced stability and 85% reduced monomeric catalytic activity, while catalytic NatA function remains unaltered. NAA10-V111G has a reduced stability compared to wild-type NAA10, and in vitro acetylation assays reveal a reduced enzymatic activity of monomeric NAA10-V111G but not for NAA10-V111G in complex with NAA15 (NatA enzymatic activity). A glycine in position 111 instead of valine will not cause any steric clashes, but loss of the more bulky hydrophobic side chain of valine may possibly cause structural alterations affecting protein stability or acetyl-CoA binding 756297 2.3.1.255 Y139A mutation in NatA, dramaitc loss of activity 736866 2.3.1.255 Y26A mutation in NatA, decrease in kcat, increase in Km value 736866 2.3.1.255 Y33A mutation in NatA, decrease in kcat, increase in Km value 736866 2.3.1.255 Y43S site-directed mutagenesis, the mutant is catalytically impaired in vitro, with approximately an 85% reduction in Nt-catalytic activity for peptide substrates EEEI, DDDI, and SESS 756675