The N-Acetylglucosamine Kinase from Yarrowia lipolytica Is a Moonlighting Protein

• Published in: International journal of molecular sciences Vol. 22; no. 23; p. 13109
• Main Authors: Flores, Carmen-Lisset, Ariño, Joaquín, Gancedo, Carlos
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 03.12.2021; MDPI
• Subjects: Arabidopsis - enzymology; Arabidopsis - genetics; Carbon; Cell walls; Cloning, Molecular; Derepression; Enzymes; Fungal Proteins - genetics; Fungal Proteins - metabolism; Gene expression; Gene Expression Profiling - methods; Gene Expression Regulation, Fungal; Genes; Genomes; Glucosamine; Glucose; Kinases; Metabolism; moonlighting; Mutation; N-acetyglucosamine kinase; N-acetyglucosamine transporter; N-Acetylglucosamine; N-Acetylglucosamine kinase; Phosphotransferases (Alcohol Group Acceptor) - genetics; Phosphotransferases (Alcohol Group Acceptor) - metabolism; Proteins; Regulation; Sequence Analysis, RNA; Transcription; Transcriptomics; Yarrowia; Yarrowia - enzymology; Yarrowia - genetics; Yarrowia - growth & development; Yarrowia lipolytica; Yeast; Yarrowia; moonlighting; N-acetyglucosamine kinase; N-acetyglucosamine transporter; yeast
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms222313109

In , expression of the genes encoding the enzymes of the N-acetylglucosamine (NAGA) utilization pathway ( genes) becomes independent of the presence of NAGA in a mutant lacking NAGA kinase. We addressed the question of whether the altered transcription was due to a lack of kinase activity or to a moonlighting role of this protein. Glucosamine-6-phosphate deaminase (Nag1) activity was measured as a reporter of genes expression. The gene encoding the NAGA transporter was deleted, creating a strain. In glucose cultures of this strain, Nag1 activity was similar to that of the strain, ruling out the possibility that NAGA derived from cell wall turnover could trigger the derepression. Heterologous NAGA kinases were expressed in a strain. Among them, the protein from did not restore kinase activity but lowered Nag1 activity 4-fold with respect to a control. Expression in the strain of YlNag5 variants F320S or D214V with low kinase activity caused a repression similar to that of the wild-type protein. Together, these results indicate that YlNag5 behaves as a moonlighting protein. An RNA-seq analysis revealed that the mutation had a limited transcriptomic effect besides derepression of the genes.