Proteomic and genomic characterization of a yeast model for Ogden syndrome [RNA-seq and Ribo-seq]

Naa10 is a Na-terminal acetyltransferase that, in a complex with its auxiliary subunit Naa15, co-translationally acetylates the a-amino group of newly synthetized proteins as they emerge from the ribosome. Roughly 40-50% of the human proteome is acetylated by Naa10, rendering this an enzyme with one of the most broad substrate ranges known. Recently, we reported an X-linked disorder of infancy, Ogden syndrome, in two families harboring a c.109T>C (p.Ser37Pro) variant in NAA10. In the present study we performed in-depth characterization of a yeast model of Ogden syndrome. Stress tests and proteomic analyses suggest that the S37P mutation disrupts Naa10 function and reduces cellular fitness during heat shock, possibly due to dysregulation of chaperone expression and accumulation. Microarray and RNA-seq revealed a pseudo-diploid gene expression profile in ?Naa10 cells, likely responsible for a mating defect. In conclusion, the data presented here further support the disruptive nature of the S37P/Ogden mutation and identify affected cellular processes potentially contributing to the severe phenotype seen in Ogden syndrome. Overall design: RNA-seq, microarray

Naa10是一种N端乙酰转移酶（N-terminal acetyltransferase），其与辅助亚基Naa15形成复合物后，可在新生蛋白从核糖体中透出时，对其α-氨基进行共翻译乙酰化修饰。人体蛋白质组中约40%~50%的蛋白可被Naa10乙酰化，使其成为已知底物范围最广泛的酶之一。此前，我们在两个携带NAA10基因c.109T>C（p.Ser37Pro）变异的家系中报道了一种婴儿期X连锁疾病——奥格登综合征（Ogden syndrome）。本研究对奥格登综合征的酵母模型进行了深度表征。应激试验与蛋白质组学分析结果显示，S37P突变会破坏Naa10的功能，并在热应激期间降低细胞适应性，这可能与分子伴侣的表达及积累异常相关。微阵列（microarray）与RNA测序（RNA-seq）分析发现，Naa10缺失（ΔNaa10）细胞呈现假二倍体基因表达谱，这大概率是其交配缺陷的成因。综上，本研究数据进一步证实了S37P/Ogden突变的破坏性，并明确了可能参与奥格登综合征严重表型的受影响细胞过程。整体实验设计：RNA测序（RNA-seq）、微阵列（microarray）