The N-acetylglucosamine kinase from Yarrowia lipolytica is a moonlighting protein
Flores, Carmen-Lisset (Universidad Autónoma de Madrid. Instituto de Investigaciones Biomédicas "Alberto Sols")
Ariño Carmona, Joaquín (Universitat Autònoma de Barcelona. Institut de Biotecnologia i de Biomedicina "Vicent Villar Palasí")
Gancedo, Carlos (Universidad Autónoma de Madrid. Instituto de Investigaciones Biomédicas "Alberto Sols")
Universitat Autònoma de Barcelona. Departament de Bioquímica i de Biologia Molecular

Data:	2021
Resum:	In Yarrowia lipolytica, expression of the genes encoding the enzymes of the N-acetylglucosamine (NAGA) utilization pathway (NAG genes) becomes independent of the presence of NAGA in a Ylnag5 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 NAG genes expression. The NGT1 gene encoding the NAGA transporter was deleted, creating a Ylnag5 ngt1 strain. In glucose cultures of this strain, Nag1 activity was similar to that of the Ylnag5 strain, ruling out the possibility that NAGA derived from cell wall turnover could trigger the derepression. Heterologous NAGA kinases were expressed in a Ylnag5 strain. Among them, the protein from Arabidopsis thaliana did not restore kinase activity but lowered Nag1 activity 4-fold with respect to a control. Expression in the Ylnag5 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 Ylnag5 mutation had a limited transcriptomic effect besides derepression of the NAG genes.
Ajuts:	Agencia Estatal de Investigación BFU2017-82574-P Agencia Estatal de Investigación PID2020-113319RB-I00
Nota:	Altres ajuts: grant CIVP18A3896 from the Fundación Ramón Areces
Drets:	Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, la comunicació pública de l'obra i la creació d'obres derivades, fins i tot amb finalitats comercials, sempre i quan es reconegui l'autoria de l'obra original.
Llengua:	Anglès
Document:	Article ; recerca ; Versió publicada
Matèria:	Moonlighting ; N-acetyglucosamine kinase ; Yarrowia ; Yeast ; N-acetyglucosamine transporter
Publicat a:	International journal of molecular sciences, Vol. 22, Issue 23 (December 2021) , art. 13109, ISSN 1422-0067

DOI: 10.3390/ijms222313109
PMID: 34884915

14 p, 7.7 MB

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