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PreprintCited by 22Plant biochemistry and biosynthesisPlant tissue culture and regenerationAlkaloids: synthesis and pharmacology

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Ancient gene clusters govern the initiation of monoterpenoid indole alkaloid biosynthesis and C3 stereochemistry inversion

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Hwang, Jaewook, Kirshner, Jonathan, Deschênes, Daniel André Ramey, Richardson, Matthew Bailey, Fleck, Steven J., Mann, Scott Galeung Alexander, Guo, Jun, Perley, Jacob Owen, Shahsavarani, Mohammadamin, Garza-Garcia, Jorge Jonathan Oswaldo, Seveck, Alyssa Dawn, Doiron, Savannah Sadie, Mai, Zhan, Silliphant, Stephen Nelson, Englehart, Sarah Anne, Blight, Barry A., Calhoun, Larry, Gao, Di, Lian, Jiazhang, Deslongchamps, Ghislain, Albert, Victor A., Qu, Yang·2025·Nature Communications
DOI 10.1101/2025.01.07.631695

Preprint — published as 10.1038/s41467-025-65543-z

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Abstract

The inversion of C3 stereochemistry in monoterpenoid indole alkaloids (MIAs), derived from the central precursor strictosidine (3 S ), is a critical step for the biosynthesis of numerous 3 R MIAs and spirooxindoles, including the antihypertensive drug reserpine. While early MIA biosynthesis preserves the 3 S configuration, the mechanism underlying C3 inversion has remained unresolved. Here, we identify and biochemically characterize a conserved oxidase-reductase pair in the Gentianales order: the heteroyohimbine/yohimbine/corynanthe C3-oxidase (HYC3O) and C3-reductase (HYC3R), which together invert the 3 S stereochemistry to 3 R across diverse substrates. Notably, HYC3O and HYC3R reside in gene clusters in Rauvolfia tetraphylla and Catharanthus roseus , homologous to an elusive geissoschizine synthase (GS) cluster we also uncovered. In R. tetraphylla , these clusters are in tandem on a single chromosome, likely derived from segmental duplication, whereas in C. roseus they reside on separate chromosomes due to translocation. Comparative genomics indicate the GS cluster originated at the base of Gentianales (∼135 Mya), coinciding with the evolution of the strictosidine synthase cluster, while the reserpine cluster arose later in rauvolfioid Apocynaceae. Together, these findings uncover the genomic and biochemical basis for key events in MIA evolution and diversification, providing insights beyond the canonical vinblastine and ajmaline biosynthetic pathways.

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Metadata

Authors
Hwang, J., Kirshner, J., Deschênes, D. A. R., Richardson, M. B., Fleck, S. J., Mann, S. G. A., Guo, J., Perley, J. O., Shahsavarani, M., Garza-Garcia, J. J. O., Seveck, A. D., Doiron, S. S., Mai, Z., Silliphant, S. N., Englehart, S. A., Blight, B. A., Calhoun, L., Gao, D., Lian, J., Deslongchamps, G., Albert, V. A., & Qu, Y.
Affiliations
  • University of New Brunswick (CA)
  • University at Buffalo, State University of New York (US)
  • Department of Chemistry, University of New Brunswick, Fredericton, E3B 5A3, Canada
  • Zhejiang University (CN)
Corresponding author
  • Deslongchamps, Ghislain
  • Albert, Victor A.
  • Qu, Yang
ORCID
Venue
Nature Communications
Publisher
openRxiv
Published
2025-01-10
Volume
16
Issue
1
Pages
10495-10495
ISSN
2041-1723 (ISSN-L)
DOI
10.1101/2025.01.07.631695
OpenAlex
W4416667963
Cited by
22
References
1
Citations by year
2026: 17 · 2025: 5
FWCI
6.58
Type
journal article
Language
en
Open access
Licence
Locations
  • Nature Communications (publishedVersion, cc-by-nc-nd)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc-nd)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
New Brunswick Innovation Foundation (RPI_2022_002, RAI_2025_041)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Plant biochemistry and biosynthesis
  • Life Sciences › Agricultural and Biological Sciences › Plant Science › Plant tissue culture and regeneration
  • Life Sciences › Pharmacology, Toxicology and Pharmaceutics › Pharmacology › Alkaloids: synthesis and pharmacology
Keywords
monoterpene indole alkaloids, strictosidine, yohimbine, biosynthetic gene clusters, reserpine, Rauvolfia tetraphylla, Catharanthus roseus, Gentianales, strictosidine synthase
MeSH terms
Multigene Family, Oxidoreductases, Phylogeny, Plant Proteins, Rauwolfia, Stereoisomerism, Vinca Alkaloids, Evolution, Molecular, Catharanthus, Secologanin Tryptamine Alkaloids, Biosynthetic Pathways

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