N-Terminal Acetyltransferase Naa40p Whereabouts Put into N-Terminal Proteoform Perspective

The evolutionary conserved N-alpha acetyltransferase Naa40p is among the most selective N-terminal acetyltransferases (NATs) identified to date. Here we identified a conserved N-terminally truncated Naa40p proteoform named Naa40p25 or short Naa40p (Naa40(S)). Intriguingly, although upon ectopic expression in yeast, both Naa40p proteoforms were capable of restoring N-terminal acetylation of the characterized yeast histone H2A Naa40p substrate, the Naa40p histone H4 substrate remained N-terminally free in human haploid cells specifically deleted for canonical Naa40p27 or 237 amino acid long Naa40p (Naa40(L)), but expressing Naa40(S). Interestingly, human Naa40(L) and Naa40(S) displayed differential expression and subcellular localization patterns by exhibiting a principal nuclear and cytoplasmic localization, respectively. Furthermore, Naa40(L) was shown to be N-terminally myristoylated and to interact with N-myristoyltransferase 1 (NMT1), implicating NMT1 in steering Naa40(L) nuclear import. Differential interactomics data obtained by biotin-dependent proximity labeling (BioID) further hints to context-dependent roles of Naa40p proteoforms. More specifically, with Naa40(S) representing the main co-translationally acting actor, the interactome of Naa40(L) was enriched for nucleolar proteins implicated in ribosome biogenesis and the assembly of ribonucleoprotein particles, overall indicating a proteoform-specific segregation of previously reported Naa40p activities. Finally, the yeast histone variant H2A.Z and the transcriptionally regulatory protein Lge1 were identified as novel Naa40p substrates, expanding the restricted substrate repertoire of Naa40p with two additional members and further confirming Lge1 as being the first redundant yNatA and yNatD substrate identified to date.

Automated summary

The study identified a novel proteoform of the evolutionary conserved N-alpha acetyltransferase Naa40p, named Naa40p25, and revealed the differences in cellular localization, translation function, and protein interactions between this proteoform and the canonical Naa40(L) form; Additionally, two new Naa40p substrates were discovered, and Lge1 was confirmed as the first redundant substrate for yNatA and yNatD.