| 2011 |
NAA60 (NatF/hNaa60) is an N-terminal acetyltransferase (NAT) with activity targeting Met-Lys- and other Met-starting protein N-termini, as demonstrated by in vitro peptide library acetylation assays using purified recombinant human and Drosophila homologues. In vivo, ectopic expression of hNaa60p in yeast acetylated distinct Met-starting yeast protein N-termini and increased general N-terminal acetylation levels. Knockdown of NAA60 in Drosophila cells induced chromosomal segregation defects. |
In vitro peptide library acetylation assay with purified recombinant protein; ectopic expression in yeast followed by N-terminal COFRADIC; RNAi knockdown in Drosophila cells |
PLoS genetics |
High |
21750686
|
| 2011 |
HAT4 (NAA60) localizes to the Golgi apparatus and functions as a B-type histone acetyltransferase that preferentially acetylates free (non-nucleosomal) histone H4 at lysine residues K79 and K91 in the globular domain. HAT4 depletion impaired nucleosome assembly, inhibited cell proliferation, sensitized cells to DNA damage, and induced apoptosis. |
Subcellular localization by immunofluorescence; in vitro acetyltransferase assay with free histones; siRNA knockdown with cell proliferation, DNA damage, and apoptosis readouts |
Molecular cell |
Medium |
21981917
|
| 2015 |
NAA60 (Naa60/NatF) localizes exclusively to the cytosolic face of Golgi membranes (established by a new membrane topology assay PROMPT and selective membrane permeabilization). NAA60 specifically acetylates N-termini of transmembrane proteins that face the cytosol, as revealed by Nt-acetylome analysis of NAA60-knockdown cells. NAA60 knockdown causes Golgi fragmentation, indicating a role in maintaining Golgi structural integrity. |
Membrane topology assay (PROMPT); selective membrane permeabilization; N-terminal acetylome (COFRADIC) analysis of knockdown cells; immunofluorescence of Golgi morphology in knockdown cells |
Cell reports |
High |
25732826
|
| 2016 |
Crystal structures of human Naa60 in complex with Ac-CoA or CoA reveal a GNAT domain followed by an amphipathic helix contributing to Golgi localization. Structural and biochemical studies identified Tyr97 and His138 as key catalytic residues, and Phe34 as influencing cofactor positioning. The non-conserved β3-β4 long loop participates in regulation of hNaa60 activity. Naa60 also harbors lysine Nε-acetyltransferase (KAT) activity toward histone H4 lysine residues. |
X-ray crystallography; site-directed mutagenesis; in vitro acetyltransferase activity assays |
Scientific reports |
High |
27550639
|
| 2017 |
The C-terminal tail of Naa60 anchors it to the Golgi as a peripheral membrane protein via two amphipathic helices that fold into α-helical conformations only in the presence of liposomes and bind membranes in a parallel orientation via hydrophobic and electrostatic interactions. Naa60 shows strong and specific binding preference for phosphatidylinositol PI(4)P-containing membranes, consistent with its primary Golgi residency. Mutational analysis confirmed the hydrophobic face of these helices is critical for membranous localization. Golgi anchoring is proposed to occur post-translationally. |
Computational modeling; in vitro liposome binding assays; cellular mutational analysis with localization readout by immunofluorescence |
The Journal of biological chemistry |
High |
28196861
|
| 2024 |
Biallelic loss-of-function variants in NAA60 cause autosomal recessive primary familial brain calcification (PFBC) with absent Nt-acetylation activity. In vitro, the phosphate importer SLC20A2 (PiT2) was identified as a direct substrate of NAA60 Nt-acetylation. In cells, NAA60 loss reduced surface levels of SLC20A2 and decreased extracellular phosphate uptake, linking NAA60-mediated Nt-acetylation of transmembrane proteins to phosphate homeostasis. |
In vitro Nt-acetylation assay with SLC20A2 as substrate; cell surface biotinylation assay; phosphate uptake assay in NAA60-deficient cells; patient variant functional analysis |
Nature communications |
High |
38480682
|
| 2024 |
A homozygous frameshift variant in NAA60 disrupts its Golgi localization and accelerates protein degradation. The mutant NAA60 alters interaction with PFBC-related proteins PiT2 (SLC20A2) and XPR1, affecting intracellular phosphate homeostasis. NAA60 KO in cells also caused decreased expression of multiple brain calcification-associated membrane proteins including RFC (reduced folate carrier), a folate metabolism protein. |
Immunofluorescence (Golgi localization); Western blot (protein stability); co-immunoprecipitation (interaction with PiT2/XPR1); mass spectrometry in NAA60 KO cell lines; gene knockout cell lines |
Movement disorders : official journal of the Movement Disorder Society |
Medium |
39229657
|
| 2025 |
BioID proximity labeling of NAA60 identified over 100 proximal partners enriched on the trans-side of the Golgi, including golgins and GRASP proteins essential for Golgi integrity. Suborganellar localization analysis revealed a more prominent medial/trans-Golgi localization of NAA60. Biotinylated peptide analysis was used to infer the topology of transmembrane protein interactors in the secretory pathway. |
BioID proximity labeling; streptavidin affinity purification; mass spectrometry; suborganellar localization analysis |
Open biology |
Medium |
39965656
|
| 2025 |
NAA60 facilitates cellular uptake of platinum-based drugs (cis- and carboplatin) by acetylating the N-termini of LRRC8A and LRRC8D (volume-regulated anion channel subunits). Loss of NAA60 decreases drug uptake and causes drug resistance. Introduction of positively charged amino acids at the N-termini of LRRC8A/D (mimicking absence of the acetyl group) similarly decreased platinum sensitivity, functionally validating the role of N-terminal acetylation at these sites. |
NAA60 knockout cells; in vitro N-terminal acetylation assay; drug uptake assay; site-directed mutagenesis of LRRC8A/D N-termini; drug sensitivity assays in BRCA1;p53-deficient cells and tumors |
Communications biology |
Medium |
41053424
|