An additional Meyerozyma guilliermondii IMH3 gene confers mycophenolic acid resistance in fungal CTG clade species.
The fungal CTG clade comprises a number of well-known yeasts that impact human health or with high biotechnological potential. To further extend the set of molecular tools dedicated to these microorganisms, the initial focus of this study was to develop a mycophenolic acid (MPA) resistance cassette. Surprisingly, while we were carrying out preliminary susceptibility testing experiments in a set of yeast species, Meyerozyma guilliermondii, although not being a MPA producer, was found to be primarily resistant toward this drug, whereas a series of nine related species were susceptible to MPA. Using comparative and functional genomic approaches, we demonstrated that all MPA-susceptible CTG clade species display a single gene, referred to as IMH3.1, encoding the MPA target inosine monophosphate dehydrogenase (IMPDH) and that MPA resistance relies on the presence in the M. guilliermondii genome of an additional IMPDH-encoding gene (IMH3.2). The M. guilliermondii IMH3.2 gene displays marked differences compared to IMH3.1 including the lack of intron, a roughly 160-fold higher transcription level and a serine residue at position 251. Placed under the control of the M. guilliermondii actin 1 gene promoter, IMH3.2 was successfully used to transform Lodderomyces elongisporus, Clavispora lusitaniae, Scheffersomyces stipitis and Candida parapsilosis.
Biomolécules et Biotechnologies Végétales, Université François-Rabelais de Tours, EA 2106, F-37200 Tours, France Groupe d'Etude des Interactions Hôte-Pathogène, Université d'Angers, EA 3142, F-49933 Angers, France.
Tatiana A
TA
Defosse
Biomolécules et Biotechnologies Végétales, Université François-Rabelais de Tours, EA 2106, F-37200 Tours, France.
Céline
C
Mélin
Biomolécules et Biotechnologies Végétales, Université François-Rabelais de Tours, EA 2106, F-37200 Tours, France.
Marc
M
Clastre
Biomolécules et Biotechnologies Végétales, Université François-Rabelais de Tours, EA 2106, F-37200 Tours, France.
Sébastien
S
Besseau
Biomolécules et Biotechnologies Végétales, Université François-Rabelais de Tours, EA 2106, F-37200 Tours, France.
Arnaud
A
Lanoue
Biomolécules et Biotechnologies Végétales, Université François-Rabelais de Tours, EA 2106, F-37200 Tours, France.
Gaëlle
G
Glévarec
Biomolécules et Biotechnologies Végétales, Université François-Rabelais de Tours, EA 2106, F-37200 Tours, France.
Audrey
A
Oudin
Biomolécules et Biotechnologies Végétales, Université François-Rabelais de Tours, EA 2106, F-37200 Tours, France.
Thomas
T
Dugé de Bernonville
Groupe d'Etude des Interactions Hôte-Pathogène, Université d'Angers, EA 3142, F-49933 Angers, France.
Patrick
P
Vandeputte
Department of Microbiology, Swedish University of Agricultural Sciences, SE-750 07 Uppsala, Sweden.
Tomas
T
Linder
Groupe d'Etude des Interactions Hôte-Pathogène, Université d'Angers, EA 3142, F-49933 Angers, France.
Jean-Philippe
JP
Bouchara
Biomolécules et Biotechnologies Végétales, Université François-Rabelais de Tours, EA 2106, F-37200 Tours, France.
Vincent
V
Courdavault
Biomolécules et Biotechnologies Végétales, Université François-Rabelais de Tours, EA 2106, F-37200 Tours, France.
Nathalie
N
Giglioli-Guivarc'h
Biomolécules et Biotechnologies Végétales, Université François-Rabelais de Tours, EA 2106, F-37200 Tours, France Groupe d'Etude des Interactions Hôte-Pathogène, Université d'Angers, EA 3142, F-49933 Angers, France nicolas.papon@univ-angers.fr.
Nicolas
N
Papon
drug resistance; fungi; mycophenolic acid; selectable marker