TY - JOUR
T1 - Molecular analysis and essentiality of Aro1 shikimate biosynthesis multi-enzyme in Candida albicans
AU - Stogios, Peter J.
AU - Liston, Sean D.
AU - Semper, Cameron
AU - Quade, Bradley
AU - Michalska, Karolina
AU - Evdokimova, Elena
AU - Ram, Shane
AU - Otwinowski, Zbyszek
AU - Borek, Dominika
AU - Cowen, Leah E.
AU - Savchenko, Alexei
N1 - Funding Information:
SD Liston was supported by a Postdoctoral Fellowship from the Natural Sciences and Engineering Research Council of Canada (516840-2018). Crystal and cryo-EM structures solved in this work were funded in whole or in part with US federal funds from the National Institute of Allergy and Infectious Diseases, National Institutes of Health, Department of Health and Human Services, under contract No. HHSN272201700060C (Center for Structural Genomics of Infectious Diseases (CSGID); http://csgid.org). LE Cowen is funded by a Canadian Institutes of Health Research (CIHR) Foundation grant (FDN-154288) and is a Canada Research Chair (Tier 1) in Microbial Genomics & Infectious Disease and co-director of the CIFAR Fungal Kingdom: Threats & Opportunities program. We thank Merck and Genome Canada for making the GRACE C. albicans mutant collection available and Andre Nantel and the National Research Council of Canada for their distribution. We thank all members of Savchenko and Cowen laboratories for their help and assistance.
Funding Information:
LE Cowen is a co-founder and shareholder in Bright Angel Therapeutics, a platform company for development of novel antifungal therapeutics. LE Cowen is a consultant for Boragen, a small-molecule development company focused on leveraging the unique chemical properties of boron chemistry for crop protection and animal health. SD Liston was supported by a Mitacs postdoctoral fellowship funded in partnership with Amplyx Pharmaceuticals Inc., now a subsidiary of Pfizer. D Borek and Z Otwinowski are co-founders of Ligo Analytics, a company that develops a software for cryogenic electron microscopy. Z Otwinowski is a co-founder of HKL Research, a company that develops software for X-ray diffraction data analysis.
Publisher Copyright:
© 2022 Stogios et al.
PY - 2022/8
Y1 - 2022/8
N2 - In the human fungal pathogen Candida albicans, ARO1 encodes an essential multi-enzyme that catalyses consecutive steps in the shikimate pathway for biosynthesis of chorismate, a precursor to folate and the aromatic amino acids. We obtained the first molecular image of C. albicans Aro1 that reveals the architecture of all five enzymatic domains and their arrangement in the context of the full-length protein. Aro1 forms a flexible dimer allowing relative autonomy of enzymatic function of the individual domains. Our activity and in cellulo data suggest that only four of Aro1's enzymatic domains are functional and essential for viability of C. albicans, whereas the 3-dehydroquinate dehydratase (DHQase) domain is inactive because of active site substitutions. We further demonstrate that in C. albicans, the type II DHQase Dqd1 can compensate for the inactive DHQase domain of Aro1, suggesting an unrecognized essential role for this enzyme in shikimate biosynthesis. In contrast, in Candida glabrata and Candida parapsilosis, which do not encode a Dqd1 homolog, Aro1 DHQase domains are enzymatically active, highlighting diversity across Candida species.
AB - In the human fungal pathogen Candida albicans, ARO1 encodes an essential multi-enzyme that catalyses consecutive steps in the shikimate pathway for biosynthesis of chorismate, a precursor to folate and the aromatic amino acids. We obtained the first molecular image of C. albicans Aro1 that reveals the architecture of all five enzymatic domains and their arrangement in the context of the full-length protein. Aro1 forms a flexible dimer allowing relative autonomy of enzymatic function of the individual domains. Our activity and in cellulo data suggest that only four of Aro1's enzymatic domains are functional and essential for viability of C. albicans, whereas the 3-dehydroquinate dehydratase (DHQase) domain is inactive because of active site substitutions. We further demonstrate that in C. albicans, the type II DHQase Dqd1 can compensate for the inactive DHQase domain of Aro1, suggesting an unrecognized essential role for this enzyme in shikimate biosynthesis. In contrast, in Candida glabrata and Candida parapsilosis, which do not encode a Dqd1 homolog, Aro1 DHQase domains are enzymatically active, highlighting diversity across Candida species.
UR - http://www.scopus.com/inward/record.url?scp=85129401244&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85129401244&partnerID=8YFLogxK
U2 - 10.26508/lsa.202101358
DO - 10.26508/lsa.202101358
M3 - Article
C2 - 35512834
AN - SCOPUS:85129401244
SN - 2575-1077
VL - 5
JO - Life Science Alliance
JF - Life Science Alliance
IS - 8
M1 - e202101358
ER -