Differentially expressed genes in Naa10p knockout mouse embryos
Ontology highlight
ABSTRACT: Mammalian Naa10p, encoded by NAA10, is the only known enzyme capable of catalyzing both N-α-acetylation of nascent proteins in cytosol and ε-acetylation of internal lysines of proteins. Both enzyme-dependent and -independent functions of Naa10p are associated with cancer and some genetic diseases with developmental defects. To examine the function of Naa10p in development, we generated Naa10-KO mice by deleting exons 2 to 6 of Naa10 gene, which encode all putative isoforms of Naa10p. RNA sequencing (RNA-seq) analysis of 2 WT and 2 Naa10p-KO embryos from the same litter was performed.
Project description:Genomic imprinting is an allelic gene expression phenomenon primarily controlled by allele-specific DNA methylation at the imprinting control region (ICR), of which the underlying mechanism remains largely unclear. Mammalian N-α-acetyltransferase 10 protein (Naa10p) catalyzes N-α-acetylation of nascent proteins. Mutation of human Naa10p is linked to severe developmental delays. Here we report that Naa10-null mice display partial embryonic lethality, growth retardation, brain disorder and maternaleffect lethality, phenotypes commonly observed in defective genomic imprinting. Genome-wide analyses further revealed global DNA hypomethylation and enriched dysregulation of imprinted genes in Naa10p-knockout embryos and ES cells. Mechanistically, Naa10p facilitates the binding of DNA methyltransferase 1 (Dnmt1) to DNA substrates and recruits Dnmt1 to ICRs of the imprinted allele during S phase. Moreover, the clinical lethal Ogden syndrome-associated mutation of Naa10p disrupts its binding to H19-ICR and Dnmt1 recruitment. Our study thus links Naa10p mutationassociated human disease to defective DNA methylation and genomic imprinting.
Project description:Genomic imprinting is an allelic gene expression phenomenon primarily controlled by allele-specific DNA methylation at the imprinting control region (ICR). How allele-specific DNA methylation at ICR is regulated is largely unknown. Mammalian N-α-acetyltransferase 10 protein (Naa10p) catalyzes N-α-acetylation of nascent proteins. Mutation of human Naa10p is linked to severe developmental defects and mental retardation, including the Ogden syndrome. Here we report that Naa10-null mice display partial embryonic lethality, growth retardation, brain disorder and maternal-effect lethality, phenotypes commonly observed in defective genomic imprinting. Genome-wide analyses reveal global DNA hypomethylation and enriched dysregulation of imprinted genes in Naa10p-null embryos and ES cells. Naa10p regulates global DNA methylation by stimulating DNA methyltransferase 1 (Dnmt1) activity, while maintaining imprinted allele methylation by binding to GCXGXG and recruits Dnmt1. Clinical mutation of Naa10p disrupts its H19-ICR binding and Dnmt1 recruitment. Our study thus links Naa10p mutation-associated human diseases to defective DNA methylation and genomic imprinting.
Project description:NAA10-mediated N-terminal acetylation is widespread and has been considered essential for viability in many model organisms. However, a Naa10 null mouse model is viable and has intact global N-terminal acetylation levels. The purpose of this project was to assess the N-terminal acetyltransferase activity of NAA15 immunoprecipitates from Naa10-KO mouse liver. We show that there is a novel paralog of Naa10 present in mice, termed Naa12, with a NatA-type enzymatic activity. We demonstrate the presence of Naa12 by detecting unique Naa12 peptides in NAA15 immunoprecipitates.
Project description:We reported that both conventional and adipose-specific Naa10p deletions in mice result in increased energy expenditure, thermogenesis, beige adipocyte differentiation and activation. Mechanistically, Naa10p acetylates the N-terminus of Pgc1α and prevents it from interacting with Ppar to activate key genes, such as Ucp1, involved in beige adipocyte function. We used microarrays to profile the gene expression changes by Naa10p KO in inguinal white adipose tissues (iWATs) derived from mice fed with high fat diet for 15 weeks.
Project description:N-alpha terminal acetylation is an abundant protein modification in eukaryotes. The NatA (N-alpha acetyltransferase A) complex is responsible for N-terminal acetylation of majority of the cytosolic proteins and its core is composed of NAA10 and NAA15 subunit. HYPK has been shown to interact with NatA core complex in human and fungus, modulating its activity. Here we address the in vivo function of the plant HYPK protein. A transcriptomics approach was applied to compare transcriptional reprogramming induced by depletion of NAA10 or HYPK mutants in plants.
Project description:N-terminal acetylation is one of the most abundant protein modifications in eukaryotes and is catalysed in humans by seven N-terminal acetyltransferases. AtNAA50 interreact with the NatA complex (NAA10 and NAA15) to form the NatE complex. In this study, we investigated the Arabidopsis thaliana N-terminal acetylome in leaf cells with KO naa50 gene to identify in vivo specific substrates using the SILProNAQ approach.
Project description:N-?-acetyltransferase 10 protein (Naa10p, also called ARD1), the catalytic subunit of N-acetyltransferase A, is a critical regulator of cell death and proliferation. Naa10p is also shown to regulate cancer metastasis by inhibiting cell motility, however its role in cancer metastasis is not fully understood. In this study, we found that high expression of Naa10p is positively correlated with the survival of patients with breast cancer, while negatively correlated with lymph node metastasisr. Naa10p inhibits breast cancer cell migration and invasion in vitro and decreases the xenograft growth and metastasis in nude mice. Microarray screening revealed that Naa10p down-regulates expression of several pro-invasive genes, which was validated by qRT-PCR analysis. To gain an insight into Naa10p’s inhibitory effect on cancer metastasis, we performed microarray analysis with RNA samples extracted from Naa10p-silenced MCF-7 (MCF-7SA) and control cells (MCF-7NC).
Project description:N-terminal protein acetylation is one of the most common protein modifications in eucaryotes and is catalyzed co-translationally by N-terminal aceytltransferases (Nats). Regarding the number of predictaed substrates, NatA is the main Nat complex in human and yeast and is composed of the catalytic subunit NAA10 and the auxilliary subunit NAA15. The down-regulation of NAA10 and NAA15 results in Arabidopsis in drought resistant plants. This study focus on the identification of altered gene expression leading to the drought resistant phenotype. Microarray analysis was used to identify misregulated genes in the NatA depleted mutants responsible for the drought resistant phenotype Arabidopsis thaliana plants were grown on soil for 6 weeks under short-day conditions (8h light / 16h dark cycles) with normal watering and subsequently challenged with drought for 10 days. After 10 days of drought Arabidopsis leaf material was harvested and used for RNA extraction and hybridization on Affymetrix microarrays. Four biological replicates from each genotype and condition were analyzed.
Project description:The female reproductive tract (FRT) is vulnerable to sexually transmitted infections and therefore a well-tuned immune surveillance system is crucial for maintaining a healthy FRT. However, our understanding of the factors that impact viral infection of the FRT and the host response are not well understood. In this work, we investigate the role of a hormonally regulated type I interferon, IFN epsilon, in control of Zika virus (ZIKV) infection of the FRT. We demonstrate that IFN epsilon has anti-ZIKV properties using a combination of IFN epsilon KO mice, blockade of endogenous IFN epsilon by neutralising Abs and rescue of IFN epsilon KO mice by recombinant IFN epsilon administered directly to the FRT.